Evaluation of mechanical and biocompatibility properties of hydroxyapatite/manganese dioxide nanocomposite scaffolds for bone tissue engineering application

The aim of this research was to evaluate the mechanical properties, biocompatibility, and degradation behavior of scaffolds made of pure hydroxyapatite (HA) and HA-modified by MnO₂ for bone tissue engineering applications. HA and MnO₂ were developed using sol-gel and precipitation methods, respectively. The scaffolds properties were characterized using X-ray diffraction (XRD), Fourier transform spectroscopy (FTIR), scanning electron microcopy (SEM), energy dispersive spectroscopy (EDS), and transmission electron microscopy (TEM). The interaction of scaffold with cells was assessed using in vitro cell proliferation and alkaline phosphatase (ALP) assays. The obtained results indicate that the HA/MnO₂ scaffolds possess higher compressive strength, toughness, hardness, and density when compared to the pure HA scaffolds. After immersing the scaffold in the SBF solution, more deposited apatite appeared on the HA/MnO₂, which results in the rougher surface on this scaffold compared to the pure HA scaffold. Finally, the in vitro biological analysis using human osteoblast cells reveals that scaffolds are biocompatible with adequate ALP activity.     

Bone morphogenetic protein‐2 immobilization on porous PCL‐BCP‐Col composite scaffolds for bone tissue engineering

The objective of this study was to develop novel porous composite scaffolds for bone tissue engineering through surface modification of polycaprolactone–biphasic calcium phosphate‐based composites (PCL–BCP). PCL–BCP composites were first fabricated with salt‐leaching method followed by aminolysis. Layer by layer (LBL) technique was then used to immobilize collagen (Col) and bone morphogenetic protein (BMP‐2) on PCL–BCP scaffolds to develop PCL–BCP–Col–BMP‐2 composite scaffold. The morphology of the composite was examined by scanning electron microscopy (SEM). The efficiency of grafting of Col and BMP‐2 on composite scaffold was measured by X‐ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). Both XPS and FTIR confirmed that Col and BMP‐2 were successfully immobilized into PCL–BCP composites. MC3TC3‐E1 preosteoblasts cells were cultivated on composites to determine the effect of Col and BMP‐2 immobilization on cell viability and proliferation. PCL–BCP–Col–BMP‐2 showed more cell attachment, cell viability, and proliferation bone factors compared to PCL–BCP‐Col composites. In addition, in vivo bone formation study using rat models showed that PCL–BCP–Col–BMP‐2 composites had better bone formation than PCL–BCP‐Col scaffold in critical size defect with 4 weeks of duration. These results suggest that PCL–BCP–Col–BMP‐2 composites can enhance bone regeneration in critical size defect in a rat model with 4 weeks of duration. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45186.     

Three-dimensional silk impregnated HAp/PHBV nanofibrous scaffolds for bone regeneration

Three-dimensional silk fibroin impregnated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofibrous scaffolds with or without hydroxyapatite (HAp) were prepared by wet-electrospinning method followed by freeze-drying. Scaffolds with cotton wool-like structure have the average fiber diameter of 450–850?nm with 80–85% porosity. In-vitro cell culture tests using MG-63 osteosarcoma human cells revealed improved cell viability, alkaline phosphatase (ALP) activity and total cellular protein amount on the silk impregnated scaffolds compared to PHBV and HAp/PHBV scaffolds after 10 days of cell culture. Immunohistochemical analyses on the silk impregnated scaffolds showed that HAp triggered cell penetration and type I collagen production. Besides, HAp mineralization tendency increased with a decrease in percent crystallinity of the scaffolds comprising HAp and silk after 4 weeks of incubation in simulated body fluid. Consequently, cotton wool-like HAp/PHBV-SF scaffold would be a promising candidate as a bone-filling material for tissue regeneration.     

Nano-hydroxyapatite and nano-hydroxyapatite/zinc oxide scaffold for bone tissue engineering application

This research aims to evaluate the mechanical properties, biocompatibility, and degradation behavior of scaffolds made of pure hydroxyapatite (HA) and HA-modified by ZnO for bone tissue engineering applications. HA and ZnO were developed using sol-gel and precipitation methods respectively. The scaffolds properties were characterized using X-ray diffraction (XRD), Fourier transform spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM), atomic absorption (AA), and atomic force microscopy (AFM). The interaction of scaffold with cells was assessed using in vitro cell proliferation and alkaline phosphatase (ALP) assays. The obtained results indicate that the HA/ZnO scaffolds possess higher compressive strength, fracture toughness, and density—but lower hardness—when compared to the pure HA scaffolds. After immersing the scaffold in the SBF solution, more deposited apatite appeared on the HA/ZnO, which results in the rougher surface on this scaffold compared to the pure HA scaffold. Finally, the in vitro biological analysis using human osteoblast cells reveals that scaffolds are biocompatible with adequate ALP activity.     

A novel bi-phase Sr-doped magnesium phosphate/calcium silicate composite scaffold and its osteogenesis promoting effect

In order to improve the osteoconductivity and the osteoinductivity of bone tissue engineering scaffold, a novel bi-phase strontium-doped magnesium phosphate/calcium silicate (Sr-MP/CSC) composite scaffold was fabricated by the self-solidifying/particulate leaching method. The bi-phase composition of the well-crystallized struvite grains wrapped by the calcium silicate floccules was propitious to the deformability and toughness of composite scaffold, and the porous structure with interconnected macropores of 100–400?µm was beneficial to supporting the tissue growth and transporting nutrients and metabolites. When the Sr-MP/CSC composite scaffolds were degraded in the simulated physiological environment, the doped strontium could be sustainably released together with Ca²⁺, Mg²⁺, PO₄^3- and silidous ions. The proliferation and osteogenic differentiation of rat bone marrow stromal cells (BMSCs) on these composite scaffolds were obviously promoted. More valuably, the Sr-doped MP/CSC scaffolds exhibited the more obvious promotion to ALP activity, Col I and OCN expression than the un-doped MP/CSC scaffold, especially in the later stage. The results suggested that the strontium combined with calcium, magnesium and silicon could synergically promote osteogenesis, and the Sr-MP/CSC composite might be one of the promising bone tissue engineering scaffold materials.     

Supercritical CO2 deposition and foaming process for fabrication of biopolyester–ZnO bone scaffolds

Subsequent supercritical CO₂‐assisted deposition and foaming process followed by in situ synthesis was used to fabricate functional polylactide (PLA) and polylactide–poly(?‐caprolactone) (PLA–PCL) bone scaffolds. Deposition of zinc bis(2‐thenoyltrifluoroacetonate) as a ZnO precursor onto biopolyester substrates (30 MPa; 110 °C) was followed by fast depressurization to create cellular structure. Contact time was optimized regarding the deposition yield (2 h), while PCL content in PLA was varied (1–10 wt %). Scaffolds impregnated with the precursor were treated with hydrazine alcoholic solution to obtain biopolyester–ZnO composites. Precursor synthesis and deposition onto the scaffolds was confirmed by Fourier‐transform infrared. Processed scaffolds had micron‐sized pores (d₅₀ ～ 20 μm). High open porosity (69–77%) and compressive strength values (2.8–8.3 MPa) corresponded to those reported for trabecular bone. PLA blending with PCL positively affected precursor deposition, crystallization rate, and compressive strength of the scaffolds. It also improved PLA surface roughness and wettability which are relevant for cell adhesion. ZnO improved compressive strength of the PLA scaffolds without significant effect on thermal stability. Analysis of structural, thermal, and mechanical properties of biopolyester–ZnO scaffolds testified a great potential of the obtained platforms as bone scaffolds. Proposed processing route is straightforward and ecofriendly, fast, easy to control, and suitable for processing of thermosensitive polymers. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 45824.     

Effects of microwave sintering on the properties of porous hydroxyapatite scaffolds

Microwave sintering was used to process porous hydroxyapatite scaffolds fabricated by the extrusion deposition technique. The effects of microwave sintering on the microstructure, phase composition, degradation, compressive strength and biological properties of the scaffolds were investigated. After rapid sintering, scaffolds with controlled structure, high densification and fine grains were obtained. A significant increase in mechanical strength was observed relative to conventional sintering. The scaffolds (55–60% porosity) microwave sintered at 1200 °C for 30 min exhibited the highest average compressive strength (45.57 MPa). The degradation was determined by immersing the scaffolds in physiological saline and monitoring the Ca²⁺concentration. The results indicated that the microwave-sintered scaffolds possessed higher solubility than conventionally sintered scaffolds, as it released more Ca²⁺ at the same temperature. Furthermore, an in vitro MC3T3-E1 cell culturing study showed significant cell adhesion, distribution, and proliferation in the microwave-sintered scaffolds. These results confirm that microwave sintering has a positive effect on the properties of porous hydroxyapatite scaffolds for bone tissue engineering applications.     

Development of Collagen/Demineralized Bone Powder Scaffolds and Periosteum-Derived Cells for Bone Tissue Engineering Application

The aim of this study was to investigate physical and biological properties of collagen (COL) and demineralized bone powder (DBP) scaffolds for bone tissue engineering. DBP was prepared and divided into three groups, based on various particle sizes: 75–125 μm, 125–250 μm, and 250–500 μm. DBP was homogeneously mixed with type I collagen and three-dimensional scaffolds were constructed, applying chemical crosslinking and lyophilization. Upon culture with human periosteum-derived cells (PD cells), osteogenic differentiation of PD cells was investigated using alkaline phosphatase (ALP) activity and calcium assay kits. The physical properties of the COL/DBP scaffolds were obviously different from COL scaffolds, irrespective of the size of DBP. In addition, PD cells cultured with COL scaffolds showed significantly higher cell adhesion and proliferation than those with COL/DBP scaffolds. In contrast, COL/DBP scaffolds exhibited greater osteoinductive potential than COL scaffolds. The PD cells with COL/DBP scaffolds possessed higher ALP activity than those with COL scaffolds. PD cells cultured with COL/DBP scaffolds with 250–500 μm particle size yielded the maximum calcium deposition. In conclusion, PD cells cultured on the scaffolds could exhibit osteoinductive potential. The composite scaffold of COL/DBP with 250–500 μm particle size could be considered a potential bone tissue engineering implant.     

Surface treatment with amino acids of porous collagen based scaffolds to improve cell adhesion and proliferation

The purpose of this study was to improve the biocompatibility of glutaraldehyde (GA) cross‐linked chitosan coated collagen scaffold for cartilage tissue regeneration. In order to prevent the potential toxicity of GA, we treated the designed scaffold with either glutamic acid or glycine. Amino acid treated scaffolds were characterized by scanning electron microscopy (SEM) techniques. Afterward, chondrocyte interaction with the composite scaffold was investigated assessing cell adhesion and proliferation using Hoechst staining and MTT cell proliferation assay, respectively. The SEM analyses of the scaffolds’ surface and cross‐section confirmed the adhesion of amino acids on the surface of the scaffolds. We also observed that scaffolds’ porosity was reduced due to the coverage of the pores by chitosan and amino acids, leading to low porosity. The use of amino acid improved the chondrocyte adhesion and proliferation inside the scaffolds’ pores when cells were cultured onto the chitosan‐coated collagen scaffolds. Overall, our in vitro results suggest the use of amino acid to improve the biocompatibility of natural polymer composite scaffold being crosslinked with glutaraldehyde. Such scaffold has improved mechanical properties; biocompatibility thus may be useful for tissue regeneration such as cartilage.

     

Osteogenic Properties of Novel Methylsulfonylmethane-Coated Hydroxyapatite Scaffold

Despite numerous advantages of using porous hydroxyapatite (HAp) scaffolds in bone regeneration, the material is limited in terms of osteoinduction. In this study, the porous scaffold made from nanosized HAp was coated with different concentrations of osteoinductive aqueous methylsulfonylmethane (MSM) solution (2.5, 5, 10, and 20%) and the corresponding MH scaffolds were referred to as MH2.5, MH5, MH10, and MH20, respectively. The results showed that all MH scaffolds resulted in burst release of MSM for up to 7 d. Cellular experiments were conducted using MC3T3-E1 preosteoblast cells, which showed no significant difference between the MH2.5 scaffold and the control with respect to the rate of cell proliferation (p > 0.05). There was no significant difference between each group at day 4 for alkaline phosphatase (ALP) activity, though the MH2.5 group showed higher level of activity than other groups at day 10. Calcium deposition, using alizarin red staining, showed that cell mineralization was significantly higher in the MH2.5 scaffold than that in the HAp scaffold (p < 0.0001). This study indicated that the MH2.5 scaffold has potential for both osteoinduction and osteoconduction in bone regeneration.     

Protein expression by bone mesenchymal stem cells cultivated in PLLA scaffolds with different pore geometry

Abstract

The effects of poly(L,L-lactide) (PLLA) scaffold with axial and isotropic structure were investigated on functional activity of rabbit bone mesenchymal stem cells (BMSCs). PLLA scaffolds were processed by freeze-dry technique at different temperatures of the scaffold frost – ?196?°C, ?25?°C and 0?°C. Scaffolds with different pore sizes were obtained by adding 5 or 10% of water phase. Scaffolds were modified by collagen type I solution. The pore sizes of polymer scaffolds were ranging from 5 to 150?µm. More protein secretion was observed in the surface-modified scaffolds than in the unmodified after 2 weeks of cultivation in vitro.     

In vitro bone engineering based on polycaprolactone and polycaprolactone–tricalcium phosphate composites

The filling of bone defects in load‐bearing areas requires scaffolds possessing physical properties that are in the range of those of the host bone. In this report, composite scaffolds comprising medical‐grade polycaprolactone and tricalcium phosphate (mPCL‐TCP) (80:20), which have been designed for load‐bearing applications, are characterized and compared with mPCL scaffolds, using in vitro studies. The composite scaffolds exhibited improved hydrophilicity, compressive modulus and strength. Human alveolar osteoblasts (AOs) grown on the composite achieved higher seeding efficiencies and more uniform distribution when compared with mPCL preparations alone. AOs demonstrated better proliferation, denser multilayered cell‐sheets and showed earlier expression of bone matrix‐related proteins on the composite than on mPCL during 28 days in vitro culture. The calcium content in the media decreased in both scaffold/cell constructs. Alkaline phosphatase activity increased significantly in mPCL matrices after osteogenic induction but no distinct change was observed in the composite. Osteocalcin expression was down‐regulated by induction in the composite but was up‐regulated in mPCL at both RNA and protein level. Immuno‐reactive signals for osteopontin and collagen type I, in combination with mineral nodules were found to be stronger in mPCL‐TCP scaffolds. We conclude that the composite scaffolds were more hydrophilic and had improved mechanical properties over mPCL scaffolds. Moreover, the primary AOs achieved better cell proliferation, and showed earlier and different matrix protein expression patterns on the composite scaffolds than on the mPCL scaffolds. Copyright © 2006 Society of Chemical Industry     

Fabrication and performance of calcium phosphate cement/small intestinal submucosa composite bionic bone scaffolds with different microstructures

The microstructure of the tissue has a very important determining effect on its performance. Herein, two calcium phosphate cement (CPC)/small intestinal submucosa(SIS) composites bionic bone scaffolds with different microstructures were fabricated by rolling or/ and assembling method. The microstructure, 3D morphology, the crystal phase and mechanical properties of the scaffolds were investigated by micro CT, XRD, FIIR, SEM and electronic universal testing machines respectively. The results showed that the pore size of all scaffolds are in the range of 100–400?µm, which are beneficial to cells growth, migration, and tissue vascularization. Their porosity and the specific surface area were 14.53?±?0.76%, 8.74?±?1.38?m²/m³ and 32?±?0.58%, 26.75?±?2.69?m²/m³ separately. The high porosity and the large specific surface area can provide a larger space and contact area for cells adhesion and proliferation. Meanwhile, compressive strength of the scaffolds soaked were 10?MPa and 27?MPa, about 1.2 folds and 3.2 folds of the original scaffolds, respectively. The results are derived from different microstructures of the scaffolds and chemical bonds between SIS and new phases (hydroxyapatite), and the scaffolds performance steadily increased at near the physiological conditions. Finally, biocompatibility of the scaffolds was evaluated by CCK8, bionic microstructure scaffolds are no cytotoxicity and their biocompatibility is favorable. Based on the microstructure, compressive strength and cytotoxicity of the scaffolds, bionic Harvarsin microstructure CPC/SIS composite scaffold is expected to turn into a scaffold with the excellent properties of real bone.     

Versatile nanostructured processing strategy for bone grafting nanocomposites based on collagen fibrillogenesis

Abstract

Strong bone-like nanocomposites of collagen type I and hydroxyapatite were prepared by coupling the in situ synthesis, hybrid gel formation and subsequent dehydration consolidation. The calcium phosphate synthesis was initiated at 4°C in the concentrated collagen monomeric solutions. Under this condition collagen molecules inhibited the calcium phosphate uncontrollably rapid growth and the predominant collagen fibril aggregation was retarded. Elevating temperature induced collagen fibrillogenesis leading to the formation of elastic hybrid gels. The dehydration consolidation of the elastic gels gave rise to strong nanocomposites. The mechanical properties and bone-like characteristics of the prepared nanocomposites were explicated. The in situ formation of a hybrid gel together with its facile processing capability suggests the versatility of this biomimetic strategy either in fabricating different structural forms (films, scaffolds and monoliths) of bone grafts or in further inclusion of other biosubstance into the nanocomposites.     

3D-printed barium strontium titanate-based piezoelectric scaffolds for bone tissue engineering

In order to promote bone healing, new generations of biomaterials are under development. These biomaterials should demonstrate proper biological and mechanical properties preferably similar to the natural bone tissue. In this research, 3D-printed barium strontium titanate (BST)/β-tricalcium phosphate (β-TCP) composite scaffolds have been synthesized as an alternative strategy for bone regeneration to not only induce appropriate bioactive characteristics but also piezoelectric behavior. The physical, chemical and biological performance of the scaffolds have been examined in terms of mechanical, dielectric properties, apatite-forming ability, Alizarin Red Staining (ARS), Alkaline Phosphatase activity (ALP), and cytotoxicity. The samples composed of 60% BST and 40% β-TCP showed the highest compressive strength, bending module, elastic modulus and the Young's modulus. The dielectric constant increased with further addition of the BST phase in the constructs. Scanning Electron Microscope (SEM) and energy dispersive X-ray (EDX) analyses showed that 60% BST/40% β-TCP sample had the highest amount of bone-like apatite formation after 28 days in simulated body fluid (SBF). Moreover, the results of ARS proved that 60% BST/40% β-TCP composite could present higher quantities of mineral deposition. The ALP activity of osteosarcoma cells on 60% BST/40% β-TCP sample showed higher activities compare with the other composites. None of the samples demonstrated any sign of toxicity using MTT test. It can be suggested that BST/β-TCP composite scaffolds can be potentially used as the next generation of bone tissue engineering scaffold materials.     

Novel β-TCP scaffold production using NaCl as a porogen for bone tissue applications

There are numerous methods for producing scaffolds to be applied in bone tissue engineering. However, the best method of scaffold production is essential to consider, with respect to their chemical composition and mechanical and structural properties, so that debris is not produced when the scaffolds are evaluated in vitro or in vivo.The primary aim of the present investigation was to produce six novel β-TCP scaffold compositions, using sodium chloride as a porogen, with two different particle sizes, measuring 1–2 mm and 750 mm-1mm, and at varied concentrations (30, 50, and 70 wt %). Physical, chemical, mechanical, and in vitro characterizations were then performed on each scaffold composition, using artificial saliva, for 7 and 14 days, with promising results. The XRD diffractograms showed the formation of two new crystalline phases (NaCaPO₄ and Ca₅[PO₄]₃Cl) in the scaffolds, after their production. In addition, scaffold porosity, Young's modulus, and the maximum resistance of compression values were in the trabecular bone range and the in vitro test, using artificial saliva, was favorable in relation to scaffold bioactivity.     

Cartilage Formation In Vivo Using High Concentration Collagen-Based Bioink with MSC and Decellularized ECM Granules

The aim of this study was to verify the applicability of high-concentration collagen-based bioink with MSC (ADSC) and decellularized ECM granules for the formation of cartilage tissue de novo after subcutaneous implantation of the scaffolds in rats. The printability of the bioink (4% collagen, 2.5% decellularized ECM granules, derived via 280 μm sieve) was shown. Three collagen-based compositions were studied: (1) with ECM; (2) with MSC; (3) with ECM and MSC. It has been established that decellularized ECM granules are able to stimulate chondrogenesis both in cell-free and MSC-laden scaffolds. Undesirable effects have been identified: bone formation as well as cartilage formation outside of the scaffold area. The key perspectives and limitations of ECM granules (powder) application have been discussed.     

Preparation and in vitro degradation of novel bioactive polylactide/wollastonite scaffolds

Composite scaffolds for applications in bone engineering from poly(D,L ‐lactide) (PDLLA) incorporated with different proportional bioactive wollastonite powders were prepared through a salt‐leaching method, using NH₄HCO₃ as porogen. The pore structures and morphology of the scaffolds were determined by scanning electron microscopy (SEM). The bioactivity of composite materials was evaluated by examining its ability to initiate the formation of hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂)(HAp) on its surface when immersed in simulated body fluids (SBF). The in vitro degradation behaviors of these scaffolds were systematically monitored at varying time periods of 1, 2, 4, 6, 8, 11, 14, 17, 20, 24, and 28 weeks postimmersion in SBF at 37°C. FT‐IR, XPS, XRD, and SEM measurements revealed that hydroxyapatite commenced to form on the surface of the scaffolds after 7 days of immersion in SBF. The measurements of weight loss, pH, and molecular weight of the samples indicated that PDLLA/wollastonite composite scaffolds degraded slower than the pure PDLLA scaffolds do. Addition of wollastonite enhanced the mechanical property of the composite scaffolds. The in vitro osteoblast culture experiment confirmed the biocompatibility of the scaffold for the growth of osteo‐blasts. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009.     

Evaluating Oxygen Tensions Related to Bone Marrow and Matrix for MSC Differentiation in 2D and 3D Biomimetic Lamellar Scaffolds

The physiological O₂ microenvironment of mesenchymal stem cells (MSCs) and osteoblasts and the dimensionality of a substrate are known to be important in regulating cell phenotype and function. By providing the physiologically normoxic environments of bone marrow (5%) and matrix (12%), we assessed their potential to maintain stemness, induce osteogenic differentiation, and enhance the material properties in the micropatterned collagen/silk fibroin scaffolds that were produced in 2D or 3D. Expression of osterix (OSX) and vascular endothelial growth factor A (VEGFA) was significantly enhanced in the 3D scaffold in all oxygen environments. At 21% O₂, OSX and VEGFA expressions in the 3D scaffold were respectively 13,200 and 270 times higher than those of the 2D scaffold. Markers for assessing stemness were significantly more pronounced on tissue culture polystyrene and 2D scaffold incubated at 5% O₂. At 21% O₂, we measured significant increases in ultimate tensile strength (p < 0.0001) and Young’s modulus (p = 0.003) of the 3D scaffold compared to the 2D scaffold, whilst 5% O₂ hindered the positive effect of cell seeding on tensile strength. In conclusion, we demonstrated that the 3D culture of MSCs in collagen/silk fibroin scaffolds provided biomimetic cues for bone progenitor cells toward differentiation and enhanced the tensile mechanical properties.     

In situ synthesized chitosan–gelatin/ZnO nanocomposite scaffold with drug delivery properties: Higher antibacterial and lower cytotoxicity effects

In this work, chitosan–gelatin/zinc oxide nanocomposite hydrogel scaffolds (CS–GEL/nZnO) were prepared via in situ synthesis of ZnO nanoparticles (nZnO) to reach a scaffold with both inherent antibacterial and drug delivery properties. The prepared nanocomposite hydrogel scaffolds were characterized using scanning electron microscopy, transmission electron microscopy, atomic absorption spectrometer, Fourier transform infrared spectroscopy, and X-ray diffraction. In addition, swelling, biodegradation, antibacterial, cytocompatibility, and cell attachment of the scaffolds were evaluated. The results showed that the prepared scaffolds had high porosity with a pore size of 50–400 μm and nZnO were well distributed without any agglomeration on the CS–GEL matrix. In addition, the nanocomposite scaffolds showed enhanced swelling, biodegradation, and antibacterial properties. Moreover, the drug delivery studies using naproxen showed that nZnO could control naproxen release. Cytocompatibility of the samples was proved using normal human dermal fibroblast cells (HFF2). In comparison to the previous reports which nZnO were simply added to the matrix of the scaffold, in situ synthesis of nZnO was led to higher antibacterial and lower cytotoxicity effects as a result of well distribution of nZnO in this method. According to the findings, the in situ synthesized CS–GEL/nZnO is strongly recommended for biomedical applications especially skin tissue engineering. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47590.