A novel scaffold for effective wound healing treatment was developed utilizing natural product bearing collagen-based biocompatible electrospun nanofibers. Initially, ostholamide (OSA) was synthesized from osthole (a natural coumarin), characterized by H,C, DEPT-135 NMR, ESI-MS, and FT-IR spectroscopy analysis. OSA was incorporated into polyhydroxybutyrate (PHB) and gelatin (GEL), which serve as templates for electrospun nanofibers. The coating of OSA-PHB-GEL nanofibers with collagen resulted in PHB-GEL-OSA-COL nanofibrous scaffold which mimics extracellular matrix and serves as an effective biomaterial for tissue engineering applications, especially for wound healing. PHB-GEL-OSA-COL, along with PHB-GEL-OSA and collagen film (COLF), was characterized in vitro and in vivo to determine its efficacy. The developed PHB-GEL-OSA-COL nanofibers posed an impressive mechanical stability, an essential requirement for wound healing. The presence of OSA had contributed to antimicrobial efficacy. These scaffolds exhibited efficient antibacterial activity against common wound pathogens, Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus). The zones of inhibition were observed to be 14 ± 22 and 10 ± 2 mm, respectively. It was observed that nanofibrous scaffold had the ability to release OSA in a controlled manner, and hence, OSA would be present at the site of application and exhibit bioactivity in a sustained manner. PHB-GEL-OSA-COL nanofiber was determined to be stable against enzymatic degradation, which is the most important parameter for promoting proliferation of cells contributing to repair and remodeling of tissues during wound healing applications. As hypothesized, PHB-GEL-OSA-COL was observed to imbibe excellent cytocompatibility, which was determined using NIH 3T3 fibroblast cell proliferation studies. PHB-GEL-OSA-COL exhibited excellent wound healing efficacy which was confirmed using full thickness excision wound model in Wistar rats. The rats treated with PHB-GEL-OSA-COL nanofibrous scaffold displayed enhanced healing when compared to untreated control. Both in vitro and in vivo analysis of PHB-GEL-OSA-COL presents a strong case of therapeutic biomaterial suiting wound repair and regeneration.

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The utilization of a multifunctional bioactive molecule functionalized electrospun dressing in tissue repair and regenerative function is a prominent therapeutic strategy for preparing efficient biomaterials to promote chronic wound healing. Designing robust and highly efficient antibacterial agents in resistance against microbes and bacterial infections is a key challenge for accelerating diabetic wound healing until today. In this study, we developed a vitamin K3 carnosine peptide (VKC)laden silk fibroin electrospun scaffold (SF-VKC) for diabetic wound healing. The structural confirmation of synthesized VKC was characterized by 1 H NMR, 13 C NMR, electrospray ionization mass spectrometry (ESI-MS), and attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy analysis, and the cell viability of VKC was evaluated by the CCK-8 assay in HFF1 and NIH 3T3 cells. VKC shows excellent cell viability on both cell lines, and the VKC and SF-VKC electrospun mats exhibited excellent antibacterial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. Prepared SF and SF-VKC fibrous mats were well characterized, and the SF-VKC nanofiber mat presented good biodegradability, adhesiveness, unique mechanical property, expedient water uptake property, sustained drug release, and excellent biocompatibility for chronic wound healing. The in vitro tissue engineering study depicted excellent cell migration and cell−cell interaction in the NIH 3T3 cells over the VKCimpregnated silk fibroin (SF-VKC) mat. A higher population of cell migration was observed in cells' denuded area (scratched region) compared to the native SF fibrous mat. Interestingly, our results demonstrated that the prepared VKC-impregnated SF mat had potentially promoted the STZ-induced diabetic wound healing in a shorter period than the pure SF mat. Thus, obtained in vitro and in vivo outcomes suggest that the VKC-laden SF electrospun fibrous mat could be a better and inexpensive fibrous antibacterial biomaterial to elicit earlier re-epithelialization and efficient matrix remodeling for accelerating chronic infected wound reconstruction in skin diabetic wound healing applications.

In this article, we describe the synthesis and biological evaluation of a novel 2-(methylamino)-3-nitro-4-(4-oxo-4Hchromen-3-yl) pyrano[3,2-c]chromen-5(4H)-one (CCN). It is also determined that CCN impregnated into the collagen scaffold has the potential to mimic the function of the extracellular matrix as a biomaterial in the field of tissue engineering. The series of pyrano[3,2-c]chromen-5(4H)-one derivatives (4a-4j), were analyzed by 1 H NMR, 13 C NMR, Mass spectral and FTIR analysis. The compound 4c confirmed by single crystal XRD studies. All the compounds were screened for antimicrobial activity against the gram positive, gram negative bacteria and yeast. Among all the compounds, the compound (4a=CCN) showed activity against Gram positive and Gram negative bacteria, when compared to the synthesized compounds. Further the compound CCN was evaluated for cytotoxicity against MCF-7, Hep-2 and Vero cancer cell lines with IC50 values of 5.4 µg/ml, 5.3 µg/ml and 68.4 µg/ml respectively. In addition, the result of flow cytometry and docking ((PDBID: 1A27 with the ligand) study were supported to the activity of the synthesized compound (4a). The FTIR and NMR analysis of the CCN impregnated collagen scaffold were done to reveal the existence of CCN molecule in the scaffold. The inherent property of the collagen scaffold was not significantly affected by structure of CCN molecule. The thermal and mechanical properties of the collagen scaffold impregnated with CCN molecule gives stability as well as supports the swelling. However, the COL-CCN scaffold showed an enhanced cell attachment and proliferation of NIH 3T3 fibroblast cells. Based on the results, the novel CCN molecule impregnated with collagen scaffold has the potential application as a biomaterial in tissue engineering.