Three-dimensional wet electrospun scaffold system for the differentiation of adipose-derived mesenchymal stem cells to islet-like clusters.

Stem cell-derived islet-like clusters (ILCs) are an alternative source of pancreatic beta cells for the treatment of diabetic mellitus. An ideal 3D culture platform for the generation of ILCs of desired cluster size is a challenge due to the clustering of islet cells in the 2D culture systems. The islet cells cultured in 2D conditions produce clusters of large size, which are less efficient in terms of insulin secretion and viability. In this study, we report that ILCs formed on a PCL-based wet electrospun fibrous scaffold with larger pore size produced clusters of the desired size, compared to that cultured on a conventional electrospun sheet. The collagen functionalization on this wet electrospun polycaprolactone (PCL) scaffold showed enhanced insulin secretion and cell viability compared to the non-functionalized or conventionally electrospun PCL scaffold. The collagen-coated wet electrospun 3D scaffold produced ILCs of cluster diameter 70 ± 20 µm and the conventionally electrospun PCL sheet produced larger ILC clusters of diameter 300 ± 10 µm. Hence the results indicate the collagen-functionalized wet electrospun scaffold system could be a potential scaffold for islet tissue engineering.

Growth and Regeneration of Osteochondral Cells in Bioactive Niche: A Promising Approach for Osteochondral Tissue Repair.

Functional repair of osteochondral defects caused due to osteoarthritis still remains the greatest challenge in orthopedic therapy. A prospective clinical strategy would be exploring osteochondral tissue engineering possibilities that promote simultaneous regeneration of the articular cartilage layer as well as the underlying subchondral bone. Incorporating the appropriate cues onto the scaffolds for the regeneration of the two contrasting tissues is therefore a demanding function. In the present study, a polymer-ceramic composite scaffolding material consisting of ternary bioactive glass (67.12 SiO2/28.5 CaO/4.38 P2O5 mol %) incorporated into a semi interpenetrating polymer network of hydrophilic-hydrophobic polymer (poly(vinyl alcohol)-polycaprolactone) matrix is prepared and physicochemically characterized. In vitro bioactivity, bone-bonding ability, and biocompatibility evaluation were performed in comparison with the pristine scaffold. The degree of chondrogenic and osteogenic potential of mesenchymal stem cells in both the scaffolds was evaluated by gene expression studies. Although both the scaffolds favored the differentiation to both cell lineages in their respective medium, a higher expression of bone specific genes found with the composite scaffold suggested that this composite scaffold would serve better for osteal layer and henceforth to promote the integration of the osteochondral construct at the defect site.

Assessing the 3D Printability of an Elastomeric Poly(caprolactone-co-lactide) Copolymer as a Potential Material for 3D Printing Tracheal Scaffolds.

The advent of 3D printing technology has made remarkable progress in the field of tissue engineering. Yet, it has been challenging to reproduce the desired mechanical properties of certain tissues by 3D printing. This was majorly due to the lack of 3D printable materials possessing mechanical properties similar to the native tissue. In this study, we have synthesized four different ratios of poly(caprolactone-co-lactide (PLCL) and tested their 3D printing capabilities. The physicochemical properties of the material were characterized using Fourier-transform infrared (FTIR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography (GPC), and differential scanning calorimetry (DSC). Furthermore, the mechanical properties were assessed using the universal testing machine (UTM). The ratio with the higher lactide content was found to have better printability. Out of the different ratios assessed, a suitable ratio having the desired mechanical properties and printability was identified and 3D printed into a tracheal scaffold. Thus, PLCL can be a potential material for 3D printing of tissues like the trachea.

Dual crosslinked pullulan-gelatin cryogel scaffold for chondrocyte-mediated cartilage repair: synthesis, characterization andin vitroevaluation.

Cryogels, a subset of hydrogels, have recently drawn attention for cartilage tissue engineering due to its inherent microporous architecture and good mechanical properties. In this study a dual crosslinked pullulan-gelatin cryogel (PDAG) scaffold was synthesized by crosslinking gelatin with oxidized pullulan by Schiff's base reaction followed by cryogelation. Chondrocytes seeded within the PDAG scaffolds and cultured for 21 din vitrodemonstrated enhanced cell proliferation, enhanced production of cartilage-specific extracellular matrix and up-regulated sulfated glycosaminoglycan without altering the articular chondrocyte phenotype. Quantitative reverse transcription-polymerase chain reaction-based gene expression studies, immunofluorescence, and histological studies demonstrated that the PDAG scaffold significantly enhanced the expression of chondrogenic marker genes such as type II collagen, aggrecan, and SOX9. Taken together, these results demonstrated that PDAG scaffold prepared by sequential Schiff's base reaction and cryogelation would be a promising cell-responsive scaffold for cartilage tissue engineering applications.

3D-printed biphasic scaffolds for the simultaneous regeneration of osteochondral tissues.

Osteochondral tissue engineering (OCTE) involves the simulation of highly complex tissues with disparate biomechanical properties. OCTE is regarded as the best option for treating osteochondral defects, most of the drawbacks of current treatment methodologies can be addressed by this method. In recent years, the conventional scaffolds used in cartilage and bone regeneration are gradually being replaced by 3D printed scaffolds (3DP). In the present study, we devised the strategy of 3D printing for fabricating biphasic and integrated scaffolds that are loaded with bioactive factors for enhancing the osteochondral tissue regeneration. Polycaprolactone (PCL) and poly(lactic-co-glycolic acid) (PLGA), is used along with bioactive factors (chondroitin sulphate and beta-tricalcium phosphate (ßTCP)) for the upper cartilage and lower bone layer respectively. The 3D printed bi-layered scaffolds with varying infill density, to mimic the native tissue, are not previously explored for OCTE. Hence, we tested the simultaneous osteochondrogenic differentiation inducing potential of the aforesaid 3D printed biphasic scaffoldsin vitro, using rabbit adipose derived mesenchymal stem cells (ADMSCs). Further, the biphasic scaffolds were highly cytocompatible, with excellent cell adhesion properties and cellular morphology. Most importantly, these biphasic scaffolds directed the simultaneous differentiation of a single stem cell population in to two cell lineages (simultaneous differentiation of rabbit ADMSCs into chondrocytes and osteoblasts). Further, these scaffolds enhanced the production of ECM and induced robust expression of marker genes that is specific for respective cartilage and bone layers. The 3D printed OCTE scaffold of our study hence can simulate the native osteochondral unit and could be potential futuristic biomimetic scaffold for osteochondral defects. Furtherin vivostudies are warranted.

Long-Term Evaluation of Allogenic Chondrocyte-Loaded PVA-PCL IPN Scaffolds for Articular Cartilage Repair in Rabbits.

OBJECTIVE: This study tested the long-term efficacy of two synthetic scaffolds for osteochondral defects and compare the outcomes with that of an established technique that uses monolayer cultured chondrocytes in a rabbit model. METHODS: Articular cartilage defect was created in both knees of 18 rabbits and divided into three groups of six in each. The defects in first group receiving cells loaded on Scaffold A (polyvinyl alcohol-polycaprolactone semi-interpenetrating polymer network (Monophasic, PVA-PCL semi-IPN), the second on Scaffold B (biphasic, PVA-PCL incorporated with bioglass as the lower layer), and the third group received chondrocytes alone. One animal from each group was sacrificed at 2 months and the rest at 1 year. O'Driscoll's score measured the quality of cartilage repair. RESULTS: The histological outcome had good scores (22, 20, and 19) for all three groups at 2 months. At 1-year follow-up, the chondrocyte alone group had the best scores (mean 20.0 ± 1.4), while the group treated by PVA-PCL semi-IPN scaffolds fared better (mean 15 ± 4.2) than the group that received biphasic scaffolds (mean 11.8 ± 5.9). In all three groups, defects treated without cells scored less than the transplant. CONCLUSION: These results indicate that while these scaffolds with chondrocytes perform well initially, their late outcome is disappointing. We propose that for all scaffold-based tissue repairs, a long-term evaluation should be mandatory. The slow degrading scaffolds need further modifications to improve the milieu for long-term growth of chondrocytes and their hyaline phenotype for the better incorporation of tissue-engineered constructs.

Keratinocytes-hair follicle bulge stem cells-fibroblasts co-cultures on a tri-layer skin equivalent derived from gelatin/PEG methacrylate nanofibers.

Generation of full thickness skin equivalent models is of increasing interest in tissue engineering because of the limitations inherent to current models. In recent years, considerable interest has been given to electrospun hybrid nanofibers prepared using natural and synthetic combinations of polymers. By blending two polymers, gelatin and PEG methacrylate we created a novel functional hydrogel-named GelMet. By adjusting the concentration of GelMet between 14 and 20wt%, three types of electrospun membranes were fabricated. Keratinocytes, hair follicle bulge stem cells (HFBSCs) and fibroblasts were successfully isolated and cultured in 14 wt%, 17 wt% and 20 wt% GelMet scaffolds respectively and generated a tri-layered electrospun construct. Characterization of GelMet electrospun membranes were compared with those of the pure gelatin nanofibers. Due to plasticity, by incorporating HFBSCs, it is expected to increase the cell content of skin substitute without the need to incorporate several different cell populations. The fiber diameter and pore size of the scaffold for each layer were fabricated in such a way to mimic the structural gradation of collagen matrix across the native skin. Good mechanical properties and dimensional stability of GelMet scaffold, combined with the ability to support cell growth in vitro, suggest its tremendous potential application in skin tissue engineering.

A nonadherent chitosan-polyvinyl alcohol absorbent wound dressing prepared via controlled freeze-dry technology.

In modern-day 21st century, the demand has increased for absorbent dressings that are nonadherent and maintain structural integrity without shedding lint in the wound site. This study looks at the development of a blend of polysaccharide chitosan and polyvinyl alcohol (PVA) and its fabrication using a novel controlled freeze-drying process, thus giving it channeled pores. The dressing was assessed for in vitro physical properties such as fluid handling, mechanical integrity, bioadhesion, and blood clotting. Additionally, cytocompatibility and hemocompatibility tests were conducted. An in vitro wound-healing assay was performed to determine the healing response. Furthermore, toxicological safety evaluation tests such as acute systemic toxicity, skin irritation, and sensitization were conducted. The results revealed that the developed dressing was biocompatible with a good absorbency rate of 0.63 ± 0.13 g/cm2, enhanced mechanical integrity, and low bioadhesive strength with good healing characteristics and nontoxic nature, which indicated that it was an ideal nonadherent absorbent wound dressing.

Tissue-engineered islet-like cell clusters generated from adipose tissue-derived stem cells on three-dimensional electrospun scaffolds can reverse diabetes in an experimental rat model and the role of porosity of scaffolds on cluster differentiation.

In the current study, three-dimensional (3D) nanofibrous scaffolds with pore sizes in the range of 24-250 µm and 24-190 µm were fabricated via a two-step electrospinning method to overcome the limitation of obtaining three-dimensionality with large pore sizes for islet culture using conventional electrospinning. The scaffolds supported the growth and differentiation of adipose-derived mesenchymal stem cells to islet-like clusters (ILCs). The pore size of the scaffolds was found to influence the cluster size, viability and insulin release of the differentiated islets. Hence, islet clusters of the desired size could be developed for transplantation to overcome the loss of bigger islets due to hypoxia which adversely impacts the outcome of transplantation. The tissue-engineered constructs with ILC diameter of 50 µm reduced glycemic value within 3-4 weeks after implantation in the omental pouch of diabetic rats. Detection of insulin in the serum of implanted rats demonstrates that the tissue-engineered construct is efficient to control hyperglycemia. Our findings prove that the 3D architecture and pore size of scaffolds regulates the morphology and size of islets during differentiation which is critical in the survival and function of ILCs in vitro and in vivo.

Glutamic acid-based dendritic peptides for scaffold-free cartilage tissue engineering.

Current treatment modalities for cartilage regeneration often result in the production of fibrous-type cartilage tissue at the defect site, which has inferior mechanical properties as compared to native hyaline cartilage. Further, effective treatments are not available at present, for preventing age-related as well as disease-related hypertrophic development of chondrocytes. In the present study, we designed and synthesized three sets of glutamic acid-based dendritic peptides, differing in degree of lipidation as well as branching. Each set constitutes of N-terminal protected as well as corresponding N-deprotected peptides. Altogether, six peptides [BE12, E12, BE3(12)4, E3(12)4, BE3OMe, E3OMe] were tested for their chondrogenesis enhancing potential in vitro, using rabbit adipose derived mesenchymal stem cells (ADMSCs). Immunohistochemical and gene expression studies as well as biochemical analyses revealed that the lipopeptides [E12 and BE3(12)4] are able to enhance chondrogenic differentiation of ADMSCs significantly (p < 0.001) as compared to control group (chondrogenic medium alone). Glycosaminoglycan content, and the chondrogenic marker genes like Aggrecan (Acan), Type II collagen (Col2a1), Hyaluronan synthase 2 (Has2), and SRY-box 9 (Sox9) expressions were found to be significantly increased in E12 and BE3(12)4 treated groups. Most importantly, the BE3(12)4 treated group showed significantly lower Type I collagen (Col1a2) and Type X collagen (Col10a1) transcript levels (p < 0.001), indicating its potential for hyaline cartilage formation and also to prevent hypertrophic development. Thus, the lipopeptides E12 and BE3(12)4 may be useful for preventing chondrocyte hypertrophy and realizing the hyaline nature of regenerated cartilage tissue in tissue engineering. STATEMENT OF SIGNIFICANCE: The current treatment modalities for degenerative cartilage diseases are unsatisfactory as the resultant regenerated cartilage is often fibrous in nature with inferior mechanical properties. Further, there is no proper treatment available for age-related development of chondrocyte hypertrophy at present. In this study we synthesized glutamic acid-based lipopeptides, which differ in the degree of lipidation as well as branching. We used a combinatorial approach of scaffold-free tissue engineering and dendritic lipopeptides to achieve hyaline-like cartilage tissue from adipose derived mesenchymal stem cells in vitro. Gene expression analysis revealed the down regulation of fibrous cartilage marker Col1a2 and hypertrophic marker Col10a1, suggesting that these lipopeptides may be useful for achieving mechanically superior hyaline cartilage regeneration in future.

Strontium functionalized scaffold for bone tissue engineering.

Drug functionalized scaffolds are currently being employed to improve local delivery of osteoprotective drugs with the aim of reducing their loading dose as well as unwanted systemic complications. In this study we tested a poly-(ε) caprolactone (PCL)-laponite-strontium ranelate (SRA) composite scaffold (PLS3) for its abilities to support growth and osteogenic differentiation of human marrow derived stromal stem cells (hMSC). The in vitro experiments showed the PLS3 scaffold supported cell growth and osteogenic differentiation. The in vivo implantation of hMSC seeded PLS3 scaffold in immunocompromised mice revealed vascularized ectopic bone formation. PLS3 scaffolds can be useful in bone regenerative applications in the fields of orthopaedics and dentistry.

Bioactive nano-fibrous scaffold for vascularized craniofacial bone regeneration.

There has been a growing demand for bone grafts for correction of bone defects in complicated fractures or tumours in the craniofacial region. Soft flexible membrane like material that could be inserted into defect by less invasive approaches; promote osteoconductivity and act as a barrier to soft tissue in growth while promoting bone formation is an attractive option for this region. Electrospinning has recently emerged as one of the most promising techniques for fabrication of extracellular matrix such as nano-fibrous scaffolds that can serve as a template for bone formation. To overcome the limitation of cell penetration of electrospun scaffolds and improve on its osteoconductive nature, in this study, we fabricated a novel electrospun composite scaffold of polyvinyl alcohol (PVA)-poly (ε) caprolactone (PCL)-Hydroxyapatite based bioceramic (HAB), namely, PVA-PCL-HAB. The scaffold prepared by dual electrospinning of PVA and PCL with HAB overcomes reduced cell attachment associated with hydrophobic PCL by combination with a hydrophilic PVA and the HAB can contribute to enhance osteoconductivity. We characterized the physicochemical and biocompatibility properties of the new scaffold material. Our results indicate PVA-PCL-HAB scaffolds support attachment and growth of stromal stem cells; [human bone marrow skeletal (mesenchymal) stem cells and dental pulp stem cells]. In addition, the scaffold supported in vitro osteogenic differentiation and in vivo vascularized bone formation. Thus, PVA-PCL-HAB scaffold is a suitable potential material for therapeutic bone regeneration in dentistry and orthopaedics.

Chitosan-agarose scaffolds supports chondrogenesis of Human Wharton's Jelly mesenchymal stem cells.

Tissue engineering strategies for cartilage aim to restore the complex biomechanical and biochemical properties of the native cartilage. To mimic the in vivo microenvironment, we developed a novel scaffold based on chitosan-agarose (CHAG scaffold) resembling the properties of native cartilage extracellular matrix (ECM) that aids in vitro cartilage formation. The CHAG scaffolds had pore size ranging from 75 to 300 µm and the degradation of 18% over 6 months in PBS. L929 cells and Human Wharton's Jelly-Mesenchymal Stem Cells (HWJ-MSCs) attached well and grew in the CHAG scaffolds. HWJ-MSCs seeded on CHAG scaffolds and cultured in chondrogenic medium were able to differentiate into chondrogenic lineage. Simultaneous supplementation of growth factors (BMP-2, TGF-ß3) significantly enhanced chondrogenesis and neo ECM synthesis. CHAG scaffolds seeded with HWJ-MSCs cultured in chondrogenic media supplemented with both BMP-2 and TGF-ß3 produced 12.71 ± 1.0 µg GAG/µg DNA compared to the one which received no or either of the growth factors. Our findings suggest that CHAG scaffolds could be used as a biomaterial scaffold for cell mediated repair approaches based on HWJ-MSCs for articular cartilage. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1845-1855, 2017.

Mechanoresponsiveness of human umbilical cord mesenchymal stem cells in in vitro chondrogenesis-A comparative study with growth factor induction.

Fetal-derived mesenchymal stem cells especially human umbilical cord matrix mesenchymal stem cells (hUCMSCs), with their ease of availability, pluripotency, and high expansion potential have emerged as an alternative solution for stem cell based cartilage therapies. An attempt to elucidate the effect of dynamic mechanical compression in modulating the chondrogenic differentiation of hUCMSCs is done in this study to add on to the knowledge of optimizing chondrogenic signals necessary for the effective differentiation of these stem cells and subsequent integration to the surrounding tissues. hUCMSCs were seeded in porous poly (vinyl alcohol)-poly (caprolactone) (PVA-PCL) scaffolds and cultured in chondrogenic medium with/without TGF-ß3 and were subjected to a dynamic compression of 10% strain, 1 Hz for 1/4 h for 7 days. The results on various analysis shows that the extent of dynamic compression is an important factor affecting cell viability. Mechanical stimulation in the form of dynamic compression stimulates expression of chondrogenic genes even in the absence of chondrogenic growth factors and also augments growth factor induced chondrogenic potential of hUCMSC. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2554-2566, 2016.

Polycaprolactone-laponite composite scaffold releasing strontium ranelate for bone tissue engineering applications.

We report polycaprolactone-laponite composite scaffold for the controlled release of strontium ranelate (SRA), a drug for osteoporosis. Laponite-SRA complex with electrostatic interaction between the drug and laponite was obtained through an aqueous phase reaction. Structural evaluation verified complexation of the bulky SRA molecules with the negatively charged laponite tactoid surfaces, leading to extended ordering of the tactoids, leaving behind the interlayer spacing of the laponite unchanged. The laponite-SRA complex was solution blended with polycaprolactone to obtain composite scaffolds. The strategy was found improving the dispersibility of laponite in PCL due to partial organomodification imparted through interaction with the SRA. The composite scaffolds with varying laponite-SRA complex content of 3-12wt% were evaluated in vitro using human osteosarcoma cells. It was confirmed that an optimum composition of the scaffold with 3wt% laponite-SRA complex loading would be ideal for obtaining enhanced ALP activity, by maintaining cell viability.

A simple and effective method for making multipotent/multilineage scaffolds with hydrophilic nature without any postmodification/treatment.

A number of biodegradable and bioresorbable materials, as well as scaffold designs, have been experimentally and/or clinically studied for tissue engineering of diverse tissue types. Cell-material responses are strongly dependent on the properties of the scaffold material. In this study, scaffolds based on polycaprolactone (PCL) and PCL blended with a triblock copolymer, Polycaprolactone-polytetrahydrofuran-polycaprolactone (PCL-PTHF-PCL) at different ratios were fabricated by electrospinning. Blending and electrospinning of the triblock copolymer with PCL generated a super hydrophilic scaffold, the mechanical and biological properties of which varied with the concentration of the triblock copolymer. The hydrophilicity of the electrospun scaffolds was determined by measurement of water-air contact angle. Cellular response to the electrospun scaffolds was studied by seeding two types of cells, L929 fibroblast cell line and rat mesenchymal stem cells (RMSC). We observed that the super hydrophilicity of the material did not prevent cell adhesion, while the cell proliferation was low or negligible for scaffolds containing higher amount of PCL-PTHF-PCL. Chondrogenic differentiation of RMSC was found to be better on the PCL blend containing 10% (w/v) of PCL-PTHF-PCL than the bare PCL. Our studies indicate that the cellular response is dependent on the biomaterial composition and highlight the importance of tailoring the scaffold properties for applications in tissue engineering and regenerative medicine.

Antibacterial nanosilver coated orthodontic bands with potential implications in dentistry.

BACKGROUND & OBJECTIVES: Fixed orthodontic treatment, an indispensable procedure in orthodontics, necessitates insertion of dental bands. Insertion of band material could also introduce a site of plaque retention. It was hypothesized that band materials with slow-release antimicrobial properties could help in sustained infection control, prevention of dental plaque formation and further associated health risks. Considering the known antimicrobial proprieties of silver, a coating of silver nanoparticle (SNP) onto the stainless steel bands was done and characterized for its beneficial properties in the prevention of plaque accumulation. METHODS: Coatings of SNPs on conventional stainless steel dental bands were prepared using thermal evaporation technology. The coated dental bands were characterized for their physicochemical properties and evaluated for antimicrobial activity and biocompatibility. The physiochemical characterization of band material both coated and uncoated was carried out using scanning electron microscope, energy dispersive spectroscopy, atomic force microscopyand contact angle test. Biocompatibility tests for coated band material were carried using L929 mouse fibroblast cell culture and MTT [3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. Antimicrobial activity of coated band material against Gram-positive bacteria was tested. RESULTS: A stable and uniform coating of SNPs was obtained. The coated band materials were biocompatible as well as possessed distinct antimicrobial activity. INTERPRETATION & CONCLUSIONS: The SNP coated dental bands could be potential antimicrobial dental bands for future clinical use. Further studies need to be done to validate the efficiency of coated band materials in oral environments.

A Novel, Single Step, Highly Sensitive In-Vitro Cell-Based Metabolic Assay Using Honeycomb Microporous Polymer Membranes.

This study describes a novel, simple and versatile system for cell-based assays at the bench-top. The system consists of Polyurethane (PU) based honeycomb membrane with the active compounds/assay reagents dispensed on its pore linings. Membranes with functionalized pores were thus created and used for conducting cell based assays. As proof-of-concept Flourocein acetate (FDA) and Propidium iodide (PI) were embedded on the pore linings and live/dead assays were performed on L929 and Hacat cell lines. The results proved the sensitivity of the membrane based cell assay. To ensure the capacity of this system for high throughput applications, membrane based live/dead assay was performed on L929 cells with varying levels of viability. The results from this experiment were quantified by microscopic and spectrofluourimetric techniques both of which were found to correlate well. It was concluded that this simple membrane based cell assay is highly versatile and enables multiple compounds to be tested on the same cell/tissue. Furthermore, this method requires low volumes of assay reagents and eliminates many of the wet techniques that are involved in a conventional assay, without compromising on the sensitivity. It is anticipated that this functionalized membrane system could be easily adapted for both manual and automated high content screening experiments including in vitro biomaterial evaluation as well as cytotoxicity of nanomaterials.

Hybrid scaffold bearing polymer-siloxane Schiff base linkage for bone tissue engineering.

Scaffolds that can provide the requisite biological cues for the fast regeneration of bone are highly relevant to the advances in tissue engineering and regenerative medicine. In the present article, we report the fabrication of a chitosan-gelatin-siloxane scaffold bearing interpolymer-siloxane Schiff base linkage, through a single-step dialdehyde cross-linking and freeze-drying method using 3-aminopropyltriethoxysilane as the siloxane precursor. Swelling of the scaffolds in phosphate buffered saline indicates enhancement with increase in siloxane concentration, whereas compressive moduli of the wet scaffolds reveal inverse dependence, owing to the presence of siloxane, rich in silanol groups. It is suggested that through the strategy of dialdehyde cross-linking, a limiting siloxane loading of 20 wt.% into a chitosan -gelatin matrix should be considered ideal for bone tissue engineering, because the scaffold made with 30 wt.% siloxane loading degrades by 48 wt.%, in 21 days. The hybrid scaffolds bearing Schiff base linkage between the polymer and siloxane, unlike the stable linkages in earlier reports, are expected to give a faster release of siloxanes and enhancement in osteogenesis. This is verified by the in vitro evaluation of the hybrid scaffolds using rabbit adipose mesenchymal stem cells, which revealed osteogenic cell-clusters on a polymer-siloxane scaffold, enhanced alkaline phosphatase activity and the expression of bone-specific genes, whereas the control scaffold without siloxane supported more of cell-proliferation than differentiation. A siloxane concentration dependent enhancement in osteogenic differentiation is also observed.