The incorporation and controlled release of platelet‐rich plasma‐derived biomolecules from polymeric tissue engineering scaffolds

Platelet‐rich plasma (PRP) has been gaining popularity in recent years as a cost‐effective material capable of stimulating healing in a number of different clinical applications. As the clinical role of PRP has been growing so too has its prevalence in the fields of tissue engineering and regenerative medicine, particularly in the field of extracellular matrix (ECM) analogue scaffold fabrication. As polymeric scaffold fabrication techniques strive to create structures that ever more closely replicate the native ECM's form and function, the need for increased scaffold bioactivity becomes more pronounced. PRP, which has been shown to contain over 300 bioactive molecules, has the potential to deliver a combination of growth factors and cytokines capable of stimulating cellular activity through enhanced chemotaxis, proliferation and ECM production. The ability to incorporate such a potent bioactive milieu into a polymeric tissue engineering scaffold, which lacks intrinsic cell signaling molecules, may help to promote scaffold integration with native tissues and increase the overall patency of polymeric ECM analogue structures. This mini‐review briefly discusses the physiological basis of PRP and its current clinical use, as well as the potential role that PRP may play in the future of polymeric tissue engineering scaffold design. Copyright © 2012 Society of Chemical Industry     

Polymeric scaffolds for cardiac tissue engineering: requirements and fabrication technologies

Cardiac tissue engineering (TE) is an emerging field, whose main goal is the development of innovative strategies for the treatment of heart diseases, with the aim of overcoming the drawbacks of traditional therapies. One of these strategies involves the implantation of three‐dimensional matrices (scaffolds) capable of supporting tissue formation. Scaffolds designed and fabricated for such application should meet several requirements, concerning both the scaffold‐forming materials and the properties of the scaffold itself. A scaffold for cardiac TE should be biocompatible and biodegradable, mimic the properties of the native cardiac tissue, provide a mechanical support to the regenerating heart and possess an interconnected porous structure to favour cell migration, nutrient and oxygen diffusion, and waste removal. Moreover, the mimesis of myocardium characteristic anisotropy is attracting increasing interest to provide engineered constructs with the possibility to be structurally and mechanically integrated in native tissue. Several conventional and non‐conventional fabrication techniques have been explored in the literature to produce polymeric scaffolds meeting all these requirements. This review describes these techniques, with a focus on their advantages and disadvantages, and their flexibility, with the final goal of providing the reader with the primal knowledge necessary to develop an effective strategy in cardiac TE. © 2013 Society of Chemical Industry     

Air‐dried 3D‐collagen–chitosan biocomposite scaffold for tissue engineering application

Collagen–chitosan scaffolds of different compositions were developed using emulsion air‐drying method. The scaffolds prepared adding 10–30 wt% of chitosan to collagen improved the mechanical properties of the composite scaffold, and 7:3 ratio (collagen :chitosan) was found to be a better composite having a tensile strength of 13.57 MPa with 9% elongation at break. The water‐uptake characteristics were performed at different pH and found to be ameliorated for the composite scaffolds compared to pure collagen and chitosan scaffold, respectively. The pores ranging from 100 to 300 μm were well interconnected, and their distribution was fairly homogeneous in the scaffold as observed through scanning electron microscopy. Furthermore, the scaffold decreased the bacterial counts and supported fibroblasts attachment and proliferation, thus demonstrating this composite to be a good substrate for biomedical application.POLYM. COMPOS., 33:2029–2035, 2012. © 2012 Society of Plastics Engineers     

Design and fabrication of novel chitin hydrogel/chitosan/nano diopside composite scaffolds for tissue engineering

In this research, novel porous composite scaffolds consisting of chitin, chitosan and nano diopside powder were prepared using the freeze-drying method. The prepared nanocomposite scaffolds were characterized by SEM, XRD, BET, TGA and FT-IR techniques. In addition, swelling, degradation and biomineralization capability, cell viability and cell attachment of the composite scaffolds were evaluated. The results indicated better swelling and degradation properties of such scaffolds their ability to become bioactive. Cytocompatibility of the scaffolds were assessed by MTT assay and cell attachment studies using Human Gingival Fibroblast cells. Cell viability studies demonstrated no sign of toxicity and cells were found to be attached to the pore walls within the scaffolds. These results suggested that the developed composite scaffolds could be a potential candidate for tissue engineering.     

Development of dual‐triggered in situ gelling scaffolds for tissue engineering

Hydrogel‐forming materials that mimic the three‐dimensional architecture and properties of tissue are known to have a positive effect on cellular differentiation and growth. A subset of those are in situ gels, which utilise in vivo conditions like pH (e.g. acetate phthalate), temperature (e.g. poloxamer) and ionic concentration (e.g. Gelrite?), and can be used to facilitate the delivery of cells to an affected tissue. Hence, we have developed in situ hydrogels based on gellan and hydroxypropylmethylcellulose (HPMC), which are known to be triggered through ions and temperature, respectively, as matrices to deliver cells. Gellan/HPMC blends had a lower gelation temperature than gellan alone crosslinked with calcium, suggesting the role of the dual trigger. Average storage modulus at a frequency of 10 Hz for gellan crosslinked with 3 mmol L^?1 calcium was 4.53 × 10³ Pa; for 9:1 gellan/HPMC crosslinked with 3 mmol L^?1 calcium was 5.59 × 10³ Pa; and for 8:2 gellan/HPMC crosslinked with 3 mmol L^?1 calcium was 2.13 × 10³ Pa, suggesting tunable stiffness by changing the gellan‐to‐HPMC ratio. Hydrophilicity was confirmed using goniometry with a contact angle much less than 90°, facilitating the passage of cells and electrolytes when using the gels as scaffolds. The gels were also found to be porous and non‐toxic to fibroblast cell line L929 and osteosarcoma cell line MG‐63, which, when encapsulated within the gels, were able to grow and proliferate. These blended hydrogels are suitable as scaffolds to encapsulate cells, with tunable stiffness modulated by varying the concentration of gellan and HPMC. © 2014 Society of Chemical Industry     

Chitin scaffolds in tissue engineering

Tissue engineering/regeneration is based on the hypothesis that healthy stem/progenitor cells either recruited or delivered to an injured site, can eventually regenerate lost or damaged tissue. Most of the researchers working in tissue engineering and regenerative technology attempt to create tissue replacements by culturing cells onto synthetic porous three-dimensional polymeric scaffolds, which is currently regarded as an ideal approach to enhance functional tissue regeneration by creating and maintaining channels that facilitate progenitor cell migration, proliferation and differentiation. The requirements that must be satisfied by such scaffolds include providing a space with the proper size, shape and porosity for tissue development and permitting cells from the surrounding tissue to migrate into the matrix. Recently, chitin scaffolds have been widely used in tissue engineering due to their non-toxic, biodegradable and biocompatible nature. The advantage of chitin as a tissue engineering biomaterial lies in that it can be easily processed into gel and scaffold forms for a variety of biomedical applications. Moreover, chitin has been shown to enhance some biological activities such as immunological, antibacterial, drug delivery and have been shown to promote better healing at a faster rate and exhibit greater compatibility with humans. This review provides an overview of the current status of tissue engineering/regenerative medicine research using chitin scaffolds for bone, cartilage and wound healing applications. We also outline the key challenges in this field and the most likely directions for future development and we hope that this review will be helpful to the researchers working in the field of tissue engineering and regenerative medicine.     

Fabrication of protein‐doped PLA composite nanofibrous scaffolds for tissue engineering

BACKGROUND: Electrospinning is known as a novel fabrication method to form nanofibrous scaffolds for tissue‐engineering application. Previously, many natural biopolymers of protein have been electrospun. However, keratin has not attracted enough attention. In this study, keratin and gelatin were co‐electrospun with polylactide (PLA), respectively. RESULTS: The resulting nanofibers were characterized by a field emission scanning electron microscope (FE‐SEM), an attenuated total reflection‐Fourier transform infrared spectroscopy (ATR‐FTIR), and an electron spectroscopy for chemical analysis (ESCA). The biodegradation of mats in the presence of trypsin solution was studied. Cell attachment experiments showed that NIH 3T3 cells adhered more and spread better onto the PLA/keratin and PLA/gelatin nanofibrous mats than that onto the blank PLA mats. MTT and BrdU assay showed that PLA/keratin and PLA/gelatin nanofibrous mats could both accelerate the viability and proliferation of fibroblast cells as compared to PLA nanofibrous mats. CONCLUSION: The present study suggests that the introduction of gelatin and keratin can both improve cell‐material interaction, especially, the former is more effective. Copyright © 2008 Society of Chemical Industry     

Tunable multifunctional tissue engineering scaffolds composed of three‐component polyampholyte polymers

Resistance to nonspecific protein adsorption and the capability to provide targeted bioactive signals are essential qualities for implantable biomaterials. The development of materials that combine these multifunctional characteristics and tunable mechanical properties has been a target in the tissue engineering field over the last decade. This study is the first to demonstrate that polyampholyte hydrogels prepared with equimolar quantities of positively charged and negatively charged monomer subunits from multiple monomer compositions have great potential to address these needs. The hydrogels were synthesized with positively charged [2‐(acryloyloxy)ethyl] trimethylammonium chloride and different monomer ratios of the negatively charged 2‐carboxyethyl acrylate and 3‐sulfopropyl methacrylate monomers. The physical and chemical properties of the hydrogels were fully characterized, including swelling, hydration, mechanical strength, and chemical composition, and the fouling resistance of the hydrogels was demonstrated using enzyme‐linked immunosorbent assays. Additionally, the capability of the hydrogels to facilitate protein conjugation via EDC/NHS conjugation chemistry was assessed. The results clearly demonstrate that the polyampholyte hydrogels have a range of tunable mechanical strength based on the monomer subunits, while maintaining their excellent nonfouling properties. Additionally, high levels of conjugated protein were achieved for all of the monomer combinations investigated. Therefore, the broadly applicable multifunctional properties of polyampholyte hydrogels and their tunable mechanical properties clearly demonstrate the potential of these materials for tissue engineering. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43985.     

Microwave processing of starch‐based porous structures for tissue engineering scaffolds

A novel microwave (MW) processing technique was used to produce biodegradable scaffolds for tissue engineering from different types of starch‐based polymers. Potato, sweet potato, corn starch, and nonisolated amaranth and quinoa starch were used to produce porous structures. Water and glycerol were used as plasticizers for the different types of starch. Characterization of the pore morphology of the scaffolds was carried out with scanning electron microscopy. Three‐dimensional structures with variable porosity and pore size distribution were obtained with the MW foaming technique. The amount of remaining water in the scaffolds and their corresponding densities showed important variations among the different types of starch. Compressive mechanical properties were assessed by indentation tests, and a strong dependence of the indentation stress on the average pore size was found. Studies in simulated body fluid were used to assess the in vitro bioactivity, degradability, and surface topology evolution in the scaffolds. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 1332–1339, 2007     

Preparation and characterization of Chitosan and PLGA‐based scaffolds for tissue engineering applications

Three dimensional (3D) biodegradable porous scaffolds play a crucial role in bone tissue repair. In this study, four types of 3D polymer/hydroxyapatite (HAp) composite scaffolds were prepared by freeze drying technique in order to mimic the organic/inorganic nature of the bone. Chitosan (CH) and poly(lactic acid‐co‐glycolic acid) (PLGA) were used as the polymeric part and HAp as the inorganic component. Properties of the resultant scaffolds, such as morphology, porosity, degradation, water uptake, mechanical and thermal stabilities were examined. 3D scaffolds having interconnected macroporous structure and 77–89% porosity were produced. The pore diameters were in the range of 6 and 200 µm. PLGA and HAp containing scaffolds had the highest compressive modulus. PLGA maintained the strength by decreasing water uptake but increased the degradation rate. Scaffolds seeded with SaOs‐2 osteoblast cells showed that all scaffolds were capable of encouraging cell adhesion and proliferation. The presence of HAp particles caused an increase in cell number on CH‐HAp scaffolds compared to CH scaffolds, while cell number decreased when PLGA was incorporated in the structure. CH‐PLGA scaffolds showed highest cell number on days 7 and 14 compared to others. Based on the properties such as interconnected porosity, high mechanical strength, and in vitro cell proliferation, blend scaffolds have the potential to be applied in hard tissue treatments. POLYM. COMPOS., 36:1917–1930, 2015. © 2014 Society of Plastics Engineers     

Collagen‐based scaffolds reinforced by chitosan fibres for bone tissue engineering

In this paper, porous bone scaffolds reinforced with chitosan fibres were prepared. The porosity and pore size of the reinforced scaffolds were both satisfactory. The reinforced scaffolds resembled natural bone in both components and crystal size. Only if the length of the fibres was no shorter than the critical length, could the fibres reinforce the material. We have proposed an empirical formula to calculate the critical length of the fibres for the porous materials and determined the modifying factor (F_l) for the porous bone scaffold investigated in this work. Along with the increase of the fibres' volume content, the compressive strength of the scaffold also increased. We have proposed a further empirical formula for calculating the compressive strength of the porous reinforced materials and determined the modifying factor (F_σ) for the porous reinforced bone scaffold examined in these studies. Along with the degradation in vitro, the decrease in strength of the reinforced scaffold was less than that of the unreinforced scaffold. The growth rate of osteoblast cells on the reinforced scaffold was higher than that on the unreinforced scaffold. These results suggest that the reinforced scaffold may be a promising candidate matrix for repairing large bone defects. Copyright © 2005 Society of Chemical Industry     

An assessment of biopolymer‐ and synthetic polymer‐based scaffolds for bone and vascular tissue engineering

The promise of tissue engineering is the combination of a scaffold with cells to initiate the regeneration of tissues or organs. Engineering of scaffolds is critical for success and tailoring of polymer properties is essential for their good performance. Many different materials of natural and synthetic origins have been investigated, but the challenge is to find those that have the right mix of mechanical performance, biodegradability and biocompatibility for biological applications. This article reviews key polymeric properties for bone and vascular scaffold eligibility with focus on biopolymers, synthetic polymers and their blends. The limitations of these polymeric systems and ways and means to improve scaffold performance specifically for bone and vascular tissue engineering are discussed. © 2013 Society of Chemical Industry     

Effects of micro and nano β‐TCP fillers in freeze‐gelled chitosan scaffolds for bone tissue engineering

Tissue engineering holds an exciting promise in providing a long‐term cure to bone‐related defects and diseases. However, one of the most important prerequisites for bone tissue engineering is an ideal platform that can aid tissue genesis by having biomimetic, mechanostable, and cytocompatible characteristics. Chitosan (CS) was chosen as the base polymer to incorporate filler, namely beta‐tri calcium phosphate (β‐TCP). This research deals with a comparative study on the properties of CS scaffolds prepared using micro‐ and nano‐sized β‐TCP as filler by freeze gelation method. The scaffolds were characterized for their morphology, porosity, swelling, structural, chemical, biodegradation, and bioresorption properties. Rheological behavior of polymer and polymer‐ceramic composite suspensions were analyzed and all the suspensions with varying ratios of β‐TCP showed non‐Newtonian behavior with shear thinning property. Pore size, porosity of micro‐ and nano‐sized composite scaffolds are measured as 48–158 μm and 77% and 43–155 μm and 81%, respectively. The scaffolds containing nano β‐TCP possess higher compressive strength (～2.67 MPa) and slower degradation rate as compared to composites prepared with micro‐sized β‐TCP (～1.52 MPa). Bioresorbability, in vitro cell viability by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay, proliferation by Alamar blue assay, cell interaction by scanning electron microscope, and florescence microscopy further validates the potentiality of freeze‐gelled CS/β‐TCP composite scaffolds for bone tissue engineering applications. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41025.     

Fabrication of co‐continuous poly(ε‐caprolactone)/polyglycolide blend scaffolds for tissue engineering

The apparent inability of a single biomaterial to meet all the requirements for tissue engineering scaffolds has led to continual research in novel engineered biomaterials. One method to provide new materials and fine‐tune their properties is via mixing materials. In this study, a biodegradable powder blend of poly(ε‐caprolactone) (PCL), polyglycolide (PGA), and poly(ethylene oxide) (PEO) was prepared and three‐dimensional interconnected porous PCL/PGA scaffolds were fabricated by combining cryomilling and compression molding/polymer leaching techniques. The resultant porous scaffolds exhibited co‐continuous morphologies with ～50% porosity. Mean pore sizes of 24 and 56 μm were achieved by varying milling time. The scaffolds displayed high mechanical properties and water uptake, in addition to a remarkably fast degradation rate. The results demonstrate the potential of this fabrication approach to obtain PCL/PGA blend scaffolds with interconnected porosity. In general, these results provide significant insight into an approach that will lead to the development of new composites and blends in scaffold manufacturing. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42471.     

Preparation and characterization of polyhydroxybutyrate‐co‐hydroxyvalerate/silk fibroin nanofibrous scaffolds for skin tissue engineering

Nanofibrous scaffolds were obtained by co‐electrospinning poly (3‐hydroxybuty‐rate‐co‐3‐hydroxyvalerate) (PHBV) and fibroin regenerated from silk in different proportions using 1,1,1,3,3,3‐hexafluoro‐2‐isopropanol (HFIP) as solvent. Field emission scanning electron microscope (FESEM) investigation showed that the fiber diameters of the nanofibrous scaffolds ranged from 190 to 460 nm. X‐ray diffraction (XRD) and Fourier transform infrared spectroscopy analysis (FT‐IR) showed that the main structure of silk fibroin (SF) in the nanofibrous scaffold was β‐sheet. Compared to the PHBV nanofibrous scaffold, the surface hydrophilicity and water‐uptake capability of the PHBV/SF nanofibrous scaffold with 50/50 were improved. The results of cell adhesion experiment showed that the fibroblasts adhered more to the PHBV/SF nanofibrous scaffold with 50/50 than the pure PHBV nanofibrous scaffold. The proliferation of fibroblast on the PHBV/SF nanofibrous scaffold with 50/50 was higher than that on the pure PHBV nanofibrous scaffold. Our results indicated that the PHBV/SF nanofibrous scaffold with 50/50 may be a better candidate for biomedical applications such as skin tissue engineering and wound dressing. POLYM. ENG. SCI., 55:907–916, 2015. © 2014 Society of Plastics Engineers     

Design and preparation of μ‐bimodal porous scaffold for tissue engineering

The aim of this study was to prepare poly‐?‐caprolactone (PCL) foams, with a well‐defined micrometric and bimodal open‐pore dimension distribution, suitable as scaffolds for tissue engineering. The porous network pathway was designed without using toxic agents by combining gas foaming (GF) and selective polymer extraction techniques. PCL was melt‐mixed with thermoplastic gelatin (TG) in concentrations ranging from 40 to 60 wt %, to achieve a cocontinuous blend morphology. The blends were subsequently gas foamed by using N₂‐CO₂ mixtures, with N₂ amount ranging from 0 to 80 vol %. Foaming temperature was changed from 38 to 110°C and different pressure drop rates were used. After foaming, TG was removed by soaking in H₂O. The effect of blend compositions and GF process parameters on foam morphologies was investigated. Results showed that different combinations of TG weight ratios and GF parameters allowed the modulation of macroporosity fraction, microporosity dimension, and degree of interconnection. By optimizing the process parameters it was possible to tailor the morphologies of highly interconnected PCL scaffolds for tissue engineering. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007     

Surface‐engineered hydroxyapatite nanocrystal/ poly(ε‐caprolactone) hybrid scaffolds for bone tissue engineering

To achieve novel polymer/bioceramic composite scaffolds for use in materials for bone tissue engineering, we prepared organic/inorganic hybrid scaffolds composed of biodegradable poly(ε‐caprolactone) (PCL) and hydroxyapatite (HA), which has excellent biocompatibility with hard tissues and high osteoconductivity and bioactivity. To improve the interactions between the scaffolds and osteoblasts, we focused on surface‐engineered, porous HA/PCL scaffolds that had HA molecules on their surfaces and within them because of the biochemical affinity between the biotin and avidin molecules. The surface modification of HA nanocrystals was performed with two different methods. Using Fourier transform infrared, X‐ray diffraction, and thermogravimetric analysis measurements, we found that surface‐modified HA nanocrystals prepared with an ethylene glycol mediated coupling method showed a higher degree of coupling (%) than those prepared via a direct coupling method. HA/PCL hybrid scaffolds with a well‐controlled porous architecture were fabricated with a gas‐blowing/particle‐leaching process. All HA/PCL scaffold samples exhibited approximately 80–85% porosity. As the HA concentration within the HA/PCL scaffolds increased, the porosity of the HA/PCL scaffolds gradually decreased. The homogeneous immobilization of biotin‐conjugated HA nanocrystals on a three‐dimensional, porous scaffold was observed with confocal microscopy. According to an in vitro cytotoxicity study, all scaffold samples exhibited greater than 80% cell viability, regardless of the HA/PCL composition or preparation method. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Integrated three‐dimensional fiber/hydrogel biphasic scaffolds for periodontal bone tissue engineering

Combining a tissue engineering scaffold made of a load‐bearing polymer with a hydrogel represents a powerful approach to enhancing the functionalities of the resulting biphasic construct, such as its mechanical properties or ability to support cellular colonization. This research activity was aimed at the development of biphasic scaffolds through the combination of an additively manufactured poly(?‐caprolactone) (PCL) fiber construct and a chitosan/poly(γ‐glutamic acid) polyelectrolyte complex hydrogel. By investigating a set of layered structures made of PCL or PCL/hydroxyapatite composite, biphasic scaffold prototypes with good integration of the two phases at the macroscale and microscale were developed. The biphasic constructs were able to absorb cell culture medium up to 10‐fold of their weight, and the combination of the two phases had a significant influence on compressive mechanical properties compared with hydrogel or PCL scaffold alone. In addition, due to the presence of chitosan in the hydrogel phase, biphasic scaffolds exerted a broad‐spectrum antibacterial activity. The developed biphasic systems appear well suited for application in periodontal bone regenerative approaches in which a biodegradable porous structure providing mechanical stability and a hydrogel phase functioning as absorbing depot of endogenous proteins are simultaneously required. © 2016 Society of Chemical Industry     

Electrospinning of poly(hydroxybutyrate‐co‐hydroxyvalerate) fibrous tissue engineering scaffolds in two different electric fields

Poly(hydroxybutyrate‐co‐hydroxyvalerate) (PHBV) was electrospun into ultrafine fibrous nonwoven mats. Different from the conventional electrospinning process, which involves a positively charged conductive needle and a grounded fiber collector (i.e., positive voltage (PV) electrospinning), pseudo‐negative voltage (NV) electrospinning, which adopted a setup such that the needle was grounded and the fiber collector was positively charged, was investigated for making ultrafine PHBV fibers. For pseudo‐NV electrospinning, the effects of various electrospinning parameters on fiber morphology and diameter were assessed systematically. The average diameters of PHBV fibers electrospun via pseudo‐NVs were compared with those of PHBV fibers electrospun via PVs. With either PV electrospinning or pseudo‐NV electrospinning, the average diameters of electrospun fibers ranged between 500 nm and 4 μm, and they could be controlled by varying the electrospinning parameters. The scientific significance and technological implication of fiber formation by PV electrospinning and pseudo‐NV electrospinning in the field of tissue engineering were discussed. POLYM. ENG. SCI., 2011. © 2011 Society of Plastics Engineers     

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.