Patent Application 15747136 - Composition For Targeted Delivery Of Nucleic

Title: Composition For Targeted Delivery Of Nucleic Acid-Based Therapeutics

Application Information

Invention Title: Composition For Targeted Delivery Of Nucleic Acid-Based Therapeutics
Application Number: 15747136
Submission Date: 2025-05-21T00:00:00.000Z
Effective Filing Date: 2018-01-23T00:00:00.000Z
Filing Date: 2018-01-23T00:00:00.000Z
National Class: 435
National Sub-Class: 325000
Examiner Employee Number: 81455
Art Unit: 1632
Tech Center: 1600

Rejection Summary

102 Rejections: 0
103 Rejections: 5

Cited Patents

The following patents were cited in the rejection:

US 8227574🔗

Office Action Text

DETAILED ACTION

The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 03/05/20/25 has been entered.
Applicant’s amendments to the claims and arguments filed on March 5, 2025 have been received and entered. Claims 1, 8, 13, and 17 have been amended. The objection to claim 13 is withdrawn in view of applicant’s amendments to claim. Claims 1, 4- 5, 8-9, 11-19 and 20 are pending in the instant application.
Election/Restrictions
Applicant’s election without traverse of species (i) in the reply filed on June 7, 2019 was acknowledged. Upon further consideration election of species requirement between different species were withdrawn and all the withdrawn species were rejoined with the elected species. The requirement was deemed proper and was therefore made FINAL.
Priority
Instant application is a 371 of PCT/US16/43762 filed on 07/22/2016 that claims priority from provisional application no 62/196,634 filed on 07/24/2015.

Information Disclosure Statement
The information disclosure statements (IDS) submitted on 03/05/2025 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement has been considered by the examiner.
Claims 1, 4- 5, 8-9, 11-19 and 20 are under consideration.

Withdrawn -Claim Rejections - 35 USC § 112
Claims 17-19 and 20 were rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. Applicant’s amendments to the claim 17 deleting the recitation of “a diameter more than 1 micron”, obviates the basis of the rejection. Applicants’ arguments with respect to the withdrawn rejections are thereby rendered moot.

New-Claim Rejections - 35 USC § 112 -necessitated by amendments
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.

The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.

Claims 13-16 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 13 is vague and indefinite to the extent the scope of phrase “tensile strength of at least 100 gF in the thread direction” could not ascertained. It is relevant to note that the recitation of “100gF (gram-Force) appears to be a unit of force and not necessarily tensile strength by itself as phrase tensile strength is a force applied per unit area. It is unclear if the 100gF is over 1nm2, 1mm2 or 1cm2 or 1m2 area of the scaffold. Claims 14-16 are included in the rejection because they directly or indirectly depend from the rejected base claim. Appropriate correction and/or clarification on record is required.

Withdrawn-Claim Rejections - 35 USC § 103
Claims 1, 4-5, 8-9, 11-14, 15 remain rejected under 35 U.S.C. 103 as being unpatentable over Pauksto et al (WO 2013/103423, dated 04/11/2013, art of record), Nie et al (Journal of Controlled Release, 2007, 120, pages 111-121, IDS) as evidenced by Baker et al (Expert Rev Med Devices. 2009 6(5): 515–532, IDS), Aneed et al (Journal of Controlled Release 94 (2004) 1-14), Behr et al (Bioconjugate Chem. 1994, 5, 382-389) further in view of Huang et al (Biomaterials, 2013, 34(16), 4038-4047, IDS) as evidenced by Nakayama et al (ACS Nano. 17 Jun 2015, Vol. 9, No. 7; pages 6900-6908, IDS). In view of Applicants’ amendment of base claim 1, introducing the limitation “a solid-state transfection”, the previous rejections of claims are hereby withdrawn. Applicants’ arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below:
Claim 13, 15 and 16 are rejected under 35 U.S.C. 103 as being unpatentable over Pauksto et al (WO 2013/103423, dated 04/11/2013, art of record), Nie et al (Journal of Controlled Release, 2007, 120, pages 111-121, IDS) as evidenced by Baker et al (Expert Rev Med Devices. 2009 6(5): 515–532, IDS), Aneed et al (Journal of Controlled Release 94 (2004) 1-14), and Behr et al (Bioconjugate Chem. 1994, 5, 382-389) , Nakayama et al (ACS Nano. 17 Jun 2015, Vol. 9, No. 7; pages 6900-6908, IDS), as applied above for claim 13 and further in view of Bolliet et al (Tissue Engineering 2008 Sep;14(3):207-19), Xu et al (The Scientific World Journal 2012, Vol. 2012; 1-10, IDS). In view of Applicants’ amendment of base claim 1, introducing the limitation “a solid-state transfection” and “a tensile strength of at least 100 gF in the thread direction”, the previous rejections of claims are hereby withdrawn. Applicants’ arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below:
Claims 17 -20 are rejected under 35 U.S.C. 103 as being unpatentable over Paukshto et al (US Patent no 8227574, dated 07/24/2012, art of record), Pauksto et al (WO 2013/103423, dated 04/11/2013, art of record), Nie et al (Journal of Controlled Release, 2007, 120, pages 111-121, IDS) Baker et al (Expert Rev Med Devices. 2009 6(5): 515–532, IDS), Aneed et al (Journal of Controlled Release 94 (2004) 1-14), Bolliet et al (Tissue Engineering 2008 Sep;14(3):207-19) and Xu et al (The Scientific World Journal 2012, Vol. 2012; 1-10, IDS). In view of Applicants’ amendment of base claim 1, introducing the limitation “a solid-state transfection”, the previous rejections of claims are hereby withdrawn. Applicants’ arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below:

New-Claim Rejections - 35 USC § 103 -necessitated by amendments
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.

Claims 1, 4-5, 8-9, 11-14, 15 and 16 rejected under 35 U.S.C. 103 as being unpatentable over Pauksto et al (WO 2013/103423, dated 04/11/2013, art of record), Nakayama et al (ACS Nano. 17 Jun 2015, Vol. 9, No. 7; pages 6900-6908, IDS) as evidenced by Huang et al (Biomaterials, 2013, 34(16), 4038-4047, IDS) and further in view of Samuel (Human Gene Therapy, 2002, 13, 791-802)/Scherer (The Journal of Gene Medicine, 2002, 634-643).
Claim interpretation: Recitation of that positively charged nanofibrillar collagen material compensate negatively charged nucleic acid molecule incorporated on the surface of the nanofibrillar collagen material is interpreted as mechanism or process of incorporation of nucleic acid on the surface of collagen nanofibrillar and facilitate transfection of the cells attached to the collagen nanofibrils by any mean including one facilitated by electrostatic interactions to bind the nucleic acid and to target cell membranes utilizing compounds selected from the group consisting of: calcium phosphate, polycations, liposomes, cationic lipids, polymers, dendrimers, nanoparticles, polyethylenimine, and polylysine (see claim 8). The term “solid state transfection” process is interpreted as nucleic acid that is immobilized on a solid surface and cells are cultured directly on that surface to facilitate uptake of the nucleic acid. Regarding recitation of tensile strength of at least 100 gF in the thread direction is indefinite for the reasons discussed above, therefore the limitation is interpreted as stress of at least 30Mpa in view the paragraph 50 of the instant specification.
With respect to claim 1, Pauksto et al teach a composition comprising aligned nanofibrillar collagen material (see para. 103) with nanoweave structure (see para. 70-74) and enabling attachment and alignment of at least one type of cells (see para 98, 100, 106); and said nanofibrillar collagen material align endothelial cells according to some embodiments of the present invention depends in part on the diameter of the collagen fibrils. It is further disclosed that variations could include nucleic acid that can enhance endothelial proliferation, maintain endothelial differentiation, and/or attract circulating endothelial progenitor cells (see para. 28). Pauksto et al discloses the modulating properties of collagen scaffolds crosslinking to increase the mechanical property and decrease degradation rate of these scaffold (para. 11 and example 2 para. 91) (limitation of claim 4). Pauksto et al teaches the tensile strength of a crosslinked collagen scaffolds to be about 25.8 MPa (see page 25).
With respect to claim 5, Pauksto et al discloses the composition further comprises a medical device comprises (figure 13 A and B, abstract).
Regarding claim 9, Pauksto et al discloses that the composition comprises cells are selected from the group consisting of myocyte precursor cells, smooth muscle cells, cardiac myocytes, skeletal myocytes, satellite cells, fibroblasts, cardiac fibroblasts, chondrocytes, osteoblasts, osteocytes, endothelial cells, epithelial cells, epidermal cells, embryonic stem cells, hemopoietic cells, neuronal cells, Schwann cells, mesenchymal stem cells, glial cells, dorsal root ganglia, anchorage-dependent cell precursors, or combinations thereof (see para. 26, claim 23).
Regarding claim 11, Pauksto discloses that the crosslinking a collagen scaffold with poly (ethylene glycol) as an example (para. 91).
With respect to claim 12, Pauksto discloses that the transfection of cells is significantly higher as compared on cells grown on the tissue culture plastic (see figure 5). Absent evidence of any unexpected superior results, it would have been obvious to one of ordinary skill in the art to optimize and select an appropriate target cell membrane utilizing compound that would facilitate the electrostatically bind the nucleic acid and to the cell type that require transfection (see figures 4 and 6C).
With respect to claim 13, Pauksto discloses a composition comprising: a thread-lik
Pauksto differs from claimed invention by not explicitly disclosing (i) thread-like nanofibrillar collagen scaffold which has a porosity of more than 80% with interconnected pores to allow capillary flow along the scaffold and exhibits mechanical strength in the thread direction at least 20 MPa and (ii) incorporating nucleic acid suggested in Pauksto is immobilized on the collagen material that modulate gene expression of the cells attached to the-nanofibrillar collagen material, such that the positively charged nanofibrillar collagen material compensate negatively charged nucleic acid incorporated on the surface of the nanofibrillar collagen material.
Nakayama provide evidence of a composition forming a thread-like nanofibrillar scaffolds with the fibrils aligned in the direction of the thread (abstract) such that the fibrils enable an attachment and alignment of at least human ECs (abstract) as in Pauksto. Nakayama further provide motivation to alter the collagen concentration, ionic strength, and the shear rate, the nanofibril diameter and pattern that could be modulated. It is further disclosed that parallel-aligned nanofibrillar sheets can be further organized into thread-like porous scaffolds, wherein these thread-like nanofibrillar scaffolds provide mechanical strength, can be surgically sutured, and can be fabricated at clinically relevant length scales (see page 6901, col. 1, para. 1) as evidenced by Huang. It is relevant to noted that Huang teaches it was routine to optimize the collagen scaffold with controlled three-dimensional nano- and micro-structure, pre-determined thickness, fibril size, and high uniformity. These scaffolds 1) better mimic the complexity of native ECM at nano- and micro-scales, 2) show high mechanical strength, 3) have uniform properties over a large area of several cm2, and 4) have controlled biodegradation rate depending on the level of crosslinking. Furthermore, these scaffolds provide a high degree of cell adhesion and confer cell guidance signals (see page 4046, col. 1, para. 1). Huang discloses the physical properties of the nanofibrils of collagen scaffold with cross-section 1.2 μm × 25000 μm (∼180 μm effective diameter) showed that its maximum load was 2.1 N in dry state, 0.9 N in wet state that amount to at least 30 MPa (stress =F/A, page 4042, col. 1para. 3). Nakayama teaches that the thread-like scaffold has porosity at least 80% with interconnected pores to allow capillary flow along the scaffold (the cross-sectional view of the fibril shows porosity and scaffold having the diameter at least 50 microns (the cross-sectional view of the fibril shows scale of more than 100 μm; Figure 1 D).

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The combination of references differs from claimed invention by not disclosing nucleic acid that is immobilized on the collagen fibril material such that nucleic acid is incorporated on the surface of the nanofibrillar collagen material.
However, before the effective filing date of instant application, it was generally known in prior art that controlled delivery of nucleic acid from collagen matrices could be achieved by regulating matrix degradation rates through implementing different crosslinking strategies and/or altering the degree of plasmid/ matrix integration. Both gene loading efficiency and retention/ release profiles have been expanded using physical crosslinking techniques such as dehydrothermal (DHT) and ultraviolet (UV) treatments, and/or chemical methods including 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatments. For instance, Samuel teaches a method to fabricate a porous gene-supplemented collagen–glycosaminoglycan (GSCG) matrix for sustained delivery (over a period of several weeks) of plasmid DNA to articular chondrocytes when implanted into cartilage lesions (see abstract). It is disclosed that Collagen–glycosaminoglycan matrices are synthesized without cross-linking, and by cross-linking treatments with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) to incorporate collagen–glycosaminoglycan matrices in solutions at different pH (see abstract) that is lyophilized. Samuel further teaches plasmid DNA continually transfected canine articular chondrocytes seeded into GSCG matrices in vitro for a 4-week period as evidenced by luciferase reporter gene expression at a higher rate as compared to cells seeded on non-crosslinked matrix (abstract, and fig. 4 and 5). It is disclosed that EDC cross linked collagen-glycosaminoglycan (GSCG) matrices supplemented with nucleic acid (luciferase-encoding genes) shows higher transgene expression in chondrocytes over a four-week period relative to non-crosslinked and DHT or UV treated matrices (see fig. 4 and 5). Samuel further contemplates the mechanism of transfection may relate to the endocytosis of solubilized plasmid DNA passively released from the collagen matrix (see page 800, col. 2, para. 3 and page 801, col. 1).
Likewise, prior art summarized by the teaching of Scherer teaches immobilizing the nucleic acid directly to the upper faces of collagen and incubated for 4 h that is subsequently lyophilized (freeze-dried). It is disclosed that collage sponges coated with naked DNA, luciferase expression was observed until day 7 for cells growing on/in the sponges and in the culture dishes (Figure 3a). Cells on PEI-DNA sponges showed gene expression throughout the experimental period of 45 days while cells in the corresponding wells expressed just for 7 days, and at a low level on day 45 (Figure 3c).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art to modify the composition of Pauksto/ Nakayama by immobilizing the nucleic acid on the surface of the aligned nanofibrils collagen, in order to provide a collagen fibril composition for gene delivery using methods disclosed in Samuel/ Scherer, as instantly claimed, with a reasonable expectation of success, before the effective filing date of the instant application. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would be motivated to do so because prior art explicitly provided motivation to use controlled three-dimensional nano- and micro-structure, pre-determined thickness, fibril size, and high uniformity, high mechanical strength and controlled biodegradation. It would have been further obvious to modify the composition of aligned nanofibrils collagen scaffold comprising nucleic acid-based molecules as taught in prior art to optimize the porosity of said scaffold that has at least 80% with interconnected pores as reported in Nakayama, in order to provide capillary flow and incorporation of nucleic acid along the scaffold, as instantly claimed, with a reasonable expectation of success. One of skill in the art would have had a reasonable expectation of success because prior art successfully reported incorporation of negatively charged nucleic acids on the surface of positively charged collagen leading to enhanced cellular uptake as exemplified by Samuel/ Scherer who reported successful ways to (i) incorporate the DNA into the cross linked collagen-glycosaminoglycan (GSCG) matrices that shows higher transgene expression in cells over a four-week period relative to non-crosslinked, or (ii) immobilize DNA on the surface of collagen fibril by simple lyophilization as in Scherer and (iii) optimize the porosity with interconnected pores to allow capillary flow along the scaffold having the diameter at least 50 microns and mechanical strength in the thread direction that is at least 30 MPa. It should be noted that the KSR case forecloses the argument that a specific teaching, suggestion, or motivation is required to support a finding of obviousness See the recent Board decision Ex parte Smith, --USPQ2d--, slip op. at 20, (Bd. Pat. App. & Interf. June 25, 2007) (citing KSR, 82 USPQ2d at 1396) ( www. uspto.gov/web/offices/dcom/bpai/prec/fd071925.pdf).

Claims 17 -20 are rejected under 35 U.S.C. 103 as being unpatentable over Paukshto et al (US Patent no 8227574, dated 07/24/2012, art of record), Pauksto et al (WO 2013/103423, dated 04/11/2013, art of record), Nakayama et al (ACS Nano. 17 Jun 2015, Vol. 9, No. 7; pages 6900-6908, IDS) as evidenced by Huang et al (Biomaterials, 2013, 34(16), 4038-4047, IDS) and further in view of Scherer (The Journal of Gene Medicine, 2002, 634-643)/or Samuel (Human Gene Therapy, 2002, 13, 791-802).
The teaching of Pauksto, Nakayama as evidenced by Huang, Scherer/Samuel have been discussed above and relied in same manner here, The combination of references differ from claimed invention by not disclosing a composition forming rod-like fibrillar micro-carrier with the micro fibrils aligned in the direction of the rod such that the fibrils.
With respect to claim 17, Paukshto (1) discloses a composition forming a rod-like fibrillar micro-carrier with the micro fibrils aligned in the direction of the rod such that the fibrils enable an attachment and alignment of at least one type of cells and human fibroblast cells are shown to attach and align on the fibrils (figures 2a-c and 10). Paukshto and Baker disclose the composition of claim 17, and Paukshto further discloses that the rod-like micro-carrier has porosity at least 80% with interconnected pores to allow capillary flow along the micro-carrier (cross sectional views of the fibril demonstrate structure; Figures 5 and 6, 13); said micro-carrier has the diameter at least 10 microns and mechanical strength in the rod direction at least 30 MPa (as further evidenced by Huang; page 6, 3rd paragraph) (limitation of claim 18). Pauksto et al (2) teach a composition comprising aligned nanofibrillar collagen material (see para. 103) with nanoweave structure (see para. 70-74) and enabling attachment and alignment of at least one type of cells (see para 98, 100, 106); and said nanofibrillar collagen material align endothelial cells according to some embodiments of the present invention depends in part on the diameter of the collagen fibrils. It is further disclosed that variations could include nucleic acid that can enhance endothelial proliferation, maintain endothelial differentiation, and/or attract circulating endothelial progenitor cells (see para. 28). Pauksto et al (1) and (2) differs from claimed invention by not explicitly disclosing incorporating nucleic acid on the nanofibrillar collagen material that modulate gene expression of the cells attached to the-nanofibrillar collagen material, such that the positively charged nanofibrillar collagen material compensate negatively charged mRNA incorporated on the surface of the nanofibrillar collagen material.
Nakayama provide evidence of a composition forming a thread-like nanofibrillar scaffolds with the fibrils aligned in the direction of the thread (abstract) such that the fibrils enable an attachment and alignment of at least human ECs (abstract) as in Pauksto. Nakayama further provide motivation to alter the collagen concentration, ionic strength, and the shear rate, the nanofibril diameter and pattern that could be modulated. It is further disclosed that parallel-aligned nanofibrillar sheets can be further organized into thread-like porous scaffolds, wherein these thread-like nanofibrillar scaffolds provide mechanical strength, can be surgically sutured, and can be fabricated at clinically relevant length scales (see page 6901, col. 1, para. 1) as evidenced by Huang. It is relevant to noted that Huang teaches it was routine to optimize the collagen scaffold with controlled three-dimensional nano- and micro-structure, pre-determined thickness, fibril size, and high uniformity. These scaffolds 1) better mimic the complexity of native ECM at nano- and micro-scales, 2) show high mechanical strength, 3) have uniform properties over a large area of several cm2, and 4) have controlled biodegradation rate depending on the level of crosslinking. Furthermore, these scaffolds provide a high degree of cell adhesion and confer cell guidance signals (see page 4046, col. 1, para. 1). Huang discloses the physical properties of the nanofibrils of collagen scaffold with cross-section 1.2 μm × 25000 μm (∼180 μm effective diameter) showed that its maximum load was 2.1 N in dry state, 0.9 N in wet state that amount to at least 30 MPa (page 4042, col. 1para. 3). Nakayama teaches that the thread-like scaffold has porosity at least 80% with interconnected pores to allow capillary flow along the scaffold (the cross-sectional view of the fibril shows porosity and scaffold having the diameter at least 50 microns (the cross-sectional view of the fibril shows scale of more than 100 μm; Figure 1 D).

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The combination of references differs from claimed invention by not disclosing nucleic acid that is immobilized on the collagen fibril material such that nucleic acid is incorporated on the surface of the nanofibrillar collagen material.
However, before the effective filing date of instant application, it was generally known in prior art that controlled delivery of nucleic acid from collagen matrices could be achieved by regulating matrix degradation rates through implementing different crosslinking strategies and/or altering the degree of plasmid/ matrix integration. Both gene loading efficiency and retention/ release profiles have been expanded using physical crosslinking techniques such as dehydrothermal (DHT) and ultraviolet (UV) treatments, and/or chemical methods including 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatments. For instance, Samuel teaches a method to fabricate a porous gene-supplemented collagen–glycosaminoglycan (GSCG) matrix for sustained delivery (over a period of several weeks) of plasmid DNA to articular chondrocytes when implanted into cartilage lesions (see abstract). It is disclosed that Collagen–glycosaminoglycan matrices are synthesized without cross-linking, and by cross-linking treatments with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) to incorporate collagen–glycosaminoglycan matrices in solutions at different pH (see abstract). Samuel further teaches plasmid DNA continually transfected canine articular chondrocytes seeded into GSCG matrices in vitro for a 4-week period as evidenced by luciferase reporter gene expression at a higher rate as compared to cells seeded on non-crosslinked matrix (abstract, and fig. 4 and 5). It is disclosed that EDC cross linked collagen-glycosaminoglycan (GSCG) matrices supplemented with nucleic acid (luciferase-encoding genes) shows higher transgene expression in chondrocytes over a four-week period relative to non-crosslinked and DHT or UV treated matrices (see fig. 4 and 5). Samuel further contemplates the mechanism of transfection may relate to the endocytosis of solubilized plasmid DNA passively released from the GSCG matrix (see page 800, col. 2, para. 3 and page 801, col. 1).
Likewise, prior art summarized by the teaching of Scherer teaches immobilizing the nucleic acid directly to the upper faces of collagen and incubated for 4 h that is subsequently lyophilized (freeze-dried). It is disclosed that collage sponges coated with naked DNA, luciferase expression was observed until day 7 for cells growing on/in the sponges and in the culture dishes (Figure 3a). Cells on PEI-DNA sponges showed gene expression throughout the experimental period of 45 days while cells in the corresponding wells expressed just for 7 days, and at a low level on day 45 (Figure 3c).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art to modify the composition comprising micro fibrils aligned in the direction of the rod such that the fibrils enable attachment and alignment of one type of cells that align on the fibrils as taught by Paukshto (1), to provide inclusion of nucleic acid based molecules as disclosed in Paukshto et al (2) and Samuel/ Scherer in order to provide a rod-like fibrillar micro-carriers aligned in the direction of the fibril that includes nuclei acid based molecules, as instantly claimed, with a reasonable expectation of success, before the effective filing date of the instant application. Said modification amounting to combining prior art elements according to known methods to yield predictable results. It would be further obvious for a person of ordinary skill in the art to modify the composition of aligned nanofibrils as taught by Pauksto, to provide tunable degradation depending on the level of crosslinking as taught by Samuel, in order for the integration of EDC/sNHS, as previously disclosed by Samuel, as instantly claimed, with a reasonable expectation of success, before the effective filing date of the instant application. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would be motivated to do so in order to provide improved flexibility in the choice of crosslink material to be used in the composition as reported in Samuel (supra). One of skill in the art would have had a reasonable expectation of success because prior art successfully reported (i) incorporation of nucleic acids on the surface of positively charged collagen leading to enhanced cellular uptake crosslinking proteins using ECD/sNHS to provide improved flexibility in the choice of crosslink material to be used in the composition as in Samuel. It should be noted that the KSR case forecloses the argument that a specific teaching, suggestion, or motivation is required to support a finding of obviousness See the recent Board decision Ex parte Smith, --USPQ2d--, slip op. at 20, (Bd. Pat. App. & Interf. June 25, 2007) (citing KSR, 82 USPQ2d at 1396) (available at http: www. uspto.gov/web/offices/dcom/bpai/prec/fd071925.pdf).

Response to arguments
To the extent that Applicants’ arguments are pertinent to the new rejections, they are addressed as follows:
Applicant’s representative disagrees with the rejection arguing amended claims disclose and cite a composition that enables a solid-state transfection where the positively charged collagen nanofibrils compensate negatively charged nucleic acid incorporated on the surface of the nanofibrillar collagen material and facilitate a solid-state transfection of the cells attached to the collagen nanofibrils. Applicants’ arguments have been fully considered, but are not found persuasive.
In response, it should be noted that the concept of sold-state transfection involving nucleic acid immobilized on a sold surface (for example a nano or micro structured substrate) and cell cultured directly on that solid surface to facilitate nucleic acid uptake was known in prior art as evident from the newly cited reference of Scherer (The Journal of Gene Medicine, 2002, 634-643) and Samuel (Human Gene Therapy, 2002, 13, 791-802). Samuel teaches immobilizing nucleic acid onto a collagen–glycosaminoglycan matrices that is synthesized by cross-linking treatments with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) in solutions at different pH (see abstract). Samuel further teaches plasmid DNA continually transfected canine articular chondrocytes seeded into GSCG matrices in vitro for a 4-week period as evidenced by luciferase reporter gene expression at a higher rate as compared to cells seeded on non-crosslinked matrix (abstract, and fig. 4 and 5). It is disclosed that EDC cross linked collagen-glycosaminoglycan (GSCG) matrices supplemented with nucleic acid (luciferase-encoding genes) show higher transgene expression in chondrocytes over a four-week period relative to non-crosslinked and DHT or UV treated matrices (see fig. 4 and 5). It is further disclosed that walls of the interconnecting pores were formed by thin strands and films of the collagen–GAG material (Fig. 1a). The average pore diameter of approximately 100 um. It is relevant to note that Samuel like instant specification discloses that nucleic acid is entrapped among the swelled collagen fibrils when the GSCG matrices are freeze-dried. Ultrastructural features consistent in appearance with plasmid could be seen by TEM in the DNA-treated collagen–GAG scaffolds but not in the untreated samples (see page 801, col. 1, para. 2). Samuel attributes the mechanism of transfection may relate to the endocytosis of solubilized plasmid DNA passively released from the GSCG matrix (see page 800, col. 2, para. 3). Therefore, it would have been prima facie obvious for a person of ordinary skill in the art to modify the composition of Pauksto/ Nakayama by immobilizing the nucleic acid in the aligned nanofibrils collagen, in order to provide a collagen fibril composition for gene delivery using methods disclosed in Samuel/ Scherer, with a reasonable expectation of success, In view of foregoing, the nucleic acid incorporated in at least one aligned nanofibrillar collagen disclosed in Paukshto et al (US Patent no 8227574)/ Pauksto et al (WO 2013/103423,) using the methods disclosed in Samuel/ Scherer would structurally be indistinguishable from the instantly recited at least one aligned nanofibrillar collagen material containing nucleic acid.
In response to applicant’s argument that there is a big difference between elastic modulus and mechanical or tensile strength as disclosed in Huang as there is no correlation between them, it should be noted that the tensile strength could be mathematically calculated dividing the maximum tensile force by the original cross-sectional area of a specimen. In this context, Huang discloses the physical properties of the nanofibrils of collagen scaffold with cross-section 1.2 μm × 25000 μm (∼180 μm effective diameter) showed that its maximum load was 2.1 N in dry state, 0.9 N in wet state that amount to at least 20 MPa (page 4042, col. 1para. 3). It would be obvious for one of ordinary skill in the art would calculate tensile strength using the formula: stress =F/A to obtain the tensile strength.
In response to applicant’s argument that neither reference teach thread like scaffold that has a porosity of more than 80% interconnected pores that exhibits the mechanical strength of at least 20 or 30 MPa, it should be noted that it is Pauksto et al who teaches a composition comprising aligned nanofibrillar collagen material (see para. 103) with nanoweave structure (see para. 70-74) and enables attachment and alignment of at least one type of cells (see para 98, 100, 106); and said nanofibrillar collagen material align endothelial cells. Pauksto et al teaches the tensile strength of a crosslinked collagen scaffolds is at least 20 MPa (see page 25, para. 95 and Huang see page 4042, col. 1, para. 2). Huang discloses the physical properties of the nanofibrils of collagen scaffold with cross-section 1.2 μm × 25000 μm (∼180 μm effective diameter) showed that its maximum load was 2.1 N in dry state, 0.9 N in wet state that amount to at least 30 MPa (stress =F/A, page 4042, col. 1para. 3). The limitation of nanofibrillar collagen scaffold having a porosity of more than 80% with interconnected pores to allow capillary flow is evident from the teaching of Nakayama. It is disclosed that aligned collagen self-assembles into a thread-like scaffold having entirely porous structure as evident from the cross-sectional view similar to figure 6 and 7 of the instant application.

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It is evident from the cross-sectional view that the scaffold is entirely porous with high capillarity similar to fig. 6 and 7 of instant application that are interconnected as required by the claim. It is further relevant note that BioBridge as disclosed in the instant specification is produced by process disclosed in US Patent no 8227574, dated 07/24/2012, art of record). Pauksto (2) teaches monolayer or multilayer stack comprising a collagen layer, wherein a surface of said collagen layer comprises: a plurality of domains with predominant orientation of rod-like fibers and a plurality of pit-like formations, , wherein said pit-like formations are filled with any living tissue cell, and encapsulated DNA (see claim 1 and 9 of ‘574). Absent evidence of any unexpected superior results, one of ordinary skill in the art reviewing the teaching of prior art would be motivated to modify and characterize the thread-like collagen scaffold as disclosed in Pauksto (1)/(2) by optimizing the porosity of said scaffold that with interconnected pores as reported in Nakayama, with reasonable expectation of success.
Therefore, in view of the fact patterns of the instant case, and the ground of rejection outlined by the examiner, applicants’ arguments on record are not compelling and do not overcome the rejection of record.

Conclusion
No claims allowed.
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure
Fang J et al 1996. Stimulation of new bone formation by direct transfer of osteogenic plasmid genes. Proceedings of the National Academy of Sciences. 93(12):5753-5758
Scherer F, Schillinger U, Putz U, Stemberger A, Plank C. 2002. Nonviral vector loaded collagen sponges for sustained gene delivery in vitro and in vivo. J. Gene Med.. 4(6):634-643
Sun et al (Biomaterials 301222–1231 (Year: 2009)) teach preparation of collagen scaffolds incorporating plasmid complexes that facilitates transfection of MSC seeded on the collagen scaffold.
Akhtar et al (Advanced Drug Delivery Reviews 59 (2007) 164–182)
Balmayor et al (USPGPUB20180214572) teach a pharmaceutical composition comprising a mRNA encoding a bone morphogenetic protein (BMP) further comprising a collagen scaffold to which said RNA has been added or into which said RNA has been loaded along with seeding increasing number of cells on collagen sponges (see para. 418, example 11, figure 17).
Shepherd et al APL Mater. 3, 014902 online 11/05/2014, 1-8.
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/ANOOP K SINGH/ Primary Examiner, Art Unit 1632