Lovastatin release from polycaprolactone coated β-tricalcium phosphate: Effects of pH,concentration and drug–polymer interactions

The approach of local drug delivery from polymeric coating is currently getting significant attention for both soft and hard tissue engineering applications for sustained and controlled release. The chemistry of the polymer and the drug, and their interactions influence the release kinetics to a great extent. Here, we examine lovastatin release behaviour from polycaprolactone (PCL) coating on β-tricalcium phosphate (β-TCP). Lovastatin was incorporated into biodegradable water insoluble PCL coating. A burst and uncontrolled lovastatin release was observed from bare β-TCP, whereas controlled and sustained release was observed from PCL coating. A higher lovastatin release was observed pH 7.4 as compared to pH 5.0. Effect of PCL concentration on lovastatin release was opposite at pH 7.4 and 5.0. At pH 5.0 lovastatin release was decreased with increasing PCL concentration, whereas release was increased with increasing PCL concentration at pH 7.4. High Ca^2 + ion concentration due to high solubility of β-TCP and degradation of PCL coating were observed at pH 5.0 compared to no detectable Ca^2 + ion release and visible degradation of PCL coating at pH 7.4. The hydrophilic–hydrophobic and hydrophobic–hydrophobic interactions between lovastatin and PCL were found to be the key factors controlling the diffusion dominated release kinetics of lovastatin from PCL coating over dissolution and degradation processes. Understanding the lovastatin release chemistry from PCL will be beneficial for designing drug delivery devices from polymeric coating or scaffolds.     

In situ growth of TiCxN1−x whiskers in Al2O3 matrix for ceramic cutting tools

For resolving the dispersing problem of whiskers and fabricating cutting tool materials with excellent properties, an in situ growth technology is used to directly synthesize TiC_xN_1?x whiskers in α-Al₂O₃ matrix by a carbothermal reduction process at a temperature range of 1250–1550 °C. The raw materials are consisted of TiO₂, carbon, nickel, and NaCl. Various molar ratios from 1:3, 1:4, 1:5 to 1:7 of TiO₂:C are experimentally used. For the molar ratio 1:3, a few whiskers can be found only at the synthesis temperature of 1550 °C. For the other molar ratios, large amount of whiskers can be observed at the whole synthesis temperature range. The highest yield of whisker is observed when the synthesis temperature is 1250 °C and the molar ratio of TiO₂:C is 1:4. The compound AlO appears at 1250 °C and AlN instead at 1550 °C. The majority of the synthesized whiskers display an ideal aspect ratio of 10–30 with a diameter of 1–3 μm. No obviously influence on the whiskers growth by the present of α-Al₂O₃ matrix powder can be noted.     

Modification of parameters in mechanochemical synthesis to obtain - and -molybdenum disilicide

In this paper, molybdenum disilicide <alpha>- and <beta>-phases can be successfully synthesized during mechanical alloying (MA). Also, this method promote a self-propagating reaction (MSR) at balls to powder ratio (BPR) 10:1, shorter milling time with speed (400 rpm) without subsequent heat treatment that was considerably lower energy than that used in conventional methods. Two different molar ratios of Mo:3Si and Mo:4Si were prepared in addition to the stoichiometric powder mixture Mo:2Si intermittent sampling was done from 4 h to 20 h. Increasing Si content clearly delayed the MSR and the reactants were gradually converted to both <alpha>- and <beta>-MoSi2 phases over a relatively long time. Samples were characterized by using X-ray diffraction (XRD)/scanning electron microscopy (SEM) analyses and grain size calculated based on the conventional Scherrer method. XRD patterns of stoichiometric powder samples milled with BPR 10:1 indicated the rapid formation of <alpha>- and <beta>-MoSi2 even after 4 h milling. Samples milled with higher BPR lost their crystallinity after milling for 16 h. SEM images in general showed considerable lowering in average particle size with milled samples. Crystallite size was found to decrease with milling time and with increasing BPR.     

Characterisation of bacterial cellulose partly acetylated by dimethylacetamide/lithium chloride

Cellulose is a water-insoluble polysaccharide used at an industrial scale for the manufacture of paper and films or in the dust form, natural, hydrolysed or derivatised. The cellulose produced by G. hansenii (former A. xylinum) has a structure identical to that of plants, but is free of lignin and hemicellulose, with several unique physical–chemical properties. The main barrier to the use of cellulose is its insolubility in water and most organic solvents, but soluble derivatives can be obtained with the use of ionic solvents. Bacterial cellulose, produced in a static, 4% glucose medium, was dissolved in hot DMAc/LiCl (120, 150 or 170 °C). The solution was analysed by ¹³C NMR, and the effect of the dissolution on the crystalline state was shown by X-ray crystallography. The crystalline structure was lost upon dissolution, becoming amorphous; this was also observed for Avicel^® plant cellulose. The soluble cellulose was partly acetylated in acetic anhydride with acetic anhydride-cellulose ratios of 1:50, 1:6 and 1:12 (w/v). The resulting cellulose acetates were examined by infrared spectroscopy, and the best result was 43% (w/v). The degree of acetylation was determined via ¹H NMR spectroscopy by comparing the area of the glucose ring at 2.60–5.20 ppm and that of the methyl proton of the acetate group at 1.80–2.20 ppm. The ¹³C NMR spectra showed acetylation at C6 ? C2 > C3 at 60–80 ppm, with C1 signals at ~ 100–104 ppm. The derivatisation of bacterial cellulose in DMAc/LiCl/acetic anhydride (1:4:50, v/v/v) gave rise to 87% substitution. The process of dissolution of the bacterial cellulose is essential for the analysis of the insoluble polymer in water, facilitating analysis and characterisation of these composites by ¹³C NMR spectroscopy, size exclusion chromatography and light scattering techniques.     

Synthesis of aligned porous poly(ε-caprolactone) (PCL)/hydroxyapatite (HA) composite microspheres

We synthesized poly(ε-caprolactone) (PCL)/hydroxyapatite (HA) composite microspheres with an aligned porous structure and evaluated their potential applications in bone tissue engineering. A range of HA particles (0, 5, 10 and 20 wt.% in relation to the PCL polymer) were added to a PCL solution in order to improve the biocompatibility of the porous PCL/HA composite microspheres. All the synthesized microspheres showed that the HA particles were distributed well in the PCL matrix, while preserving their aligned porous structure. The average size of the PCL/HA composite microspheres increased from 62 ± 7 to 179 ± 95 μm with increasing HA content from 0 to 20 wt.%. The incorporation of the HA particles to the PCL polymer led to a considerable improvement in in vitro bioactivity, which was assessed by immersing the PCL/HA composite microspheres in simulated body fluid (SBF). A number of apatite crystals could be precipitated on the surface of the aligned porous PCL/HA composite microspheres after soaking in the SBF for 7 days.     

Sol–gel derived nanoscale bioactive glass (NBG) particles reinforced poly(ε-caprolactone) composites for bone tissue engineering

This study investigated the effect of the addition of sol–gel derived nanoscale bioactive glass (NBG) particles on the mechanical properties and biological performances of PCL polymer, in order to evaluate the potential applications of PCL/NBG composites for bone tissue regeneration. Regardless of the NBG contents (10, 20, and 30 wt.%), the NBG particles, which were synthesized through the sol–gel process using polyethylene glycol (PEG) polymer as a template, could be uniformly dispersed in the PCL matrix, while generating pores in the PCL/NBG composites. The elastic modulus of the PCL/NBG composites increased remarkably from 89 ± 11 MPa to 383 ± 50 MPa with increasing NBG content from 0 to 30 wt.%, while still showing good ultimate tensile strength in the range of 15–19 MPa. The hydrophilicity, water absorption and degradation behavior of the PCL/NBG composites were also enhanced by the addition of the NBG particles. Furthermore, the PCL/NBG composite with a NBG content of 30 wt.% showed significantly enhanced in vitro bioactivity and cellular response compared to those of the pure PCL.     

Tricalcium phosphate and tricalcium phosphate/polycaprolactone particulate composite for controlled release of protein

β-Tricalcium phosphate (β-TCP) with three different particle size ranges was used to study the effects of particle size and surface area on protein adsorption and release. Polycaprolactone (PCL) coating was applied on the particle systems to investigate its effect on particulate system properties from both structural and application aspects. The maximum loading of 27 mg/g was achieved for 100 nm particles. Bovine serum albumin (BSA) loading amount was controlled by varying the BSA loading solution concentration, as well as the sample powder's surface area. Increasing the surface area of the delivery powder significantly increased loading and release yield. Unlike the samples with low surface area, the lowest particle size samples showed sigmoidal release profile. This indicated that release was governed by different mechanisms for particles with different sizes. While the majority of samples showed no more than 50% release, the 550 nm particles demonstrated 100% release. PCL coating showed no significant ability to attenuate burst release in PBS. However, it led to a steadier release profile as compared to the bare TCP particles. FTIR analysis also proved that the secondary structure of BSA did not change significantly during the adsorption; however, minor denaturation was found during the release. The same results were found when PCL coating was applied on the TCP particles. We envision potential use of TCP and TCP + PCL systems in bone growth factor or orthopedic drug delivery applications in future bone tissue engineering application.     

Polymerization of a water soluble glucose vinyl ester monomer with tensoactive properties synthesized by enzymatic catalyst

The aim of this study was to synthesize a reducing sugar branched polymer, using d-glucose as carbohydrate and divinyl adipate (DVA) as vegetable oil derivative, and to use it as an environmentally-friendly material. Several reaction conditions were examined in order to reach high conversion rates during product formation. The main parameters evaluated were: type of protease (neutral and alkaline) used as catalyst, reaction temperature (30 °C, 50 °C and 70 °C), sugar/DVA molar ratio (1/1, 1/2, 1/4) and water/DMF content (0%, 1%, 3%, 5%, 10%, 20% (v/v)). After choosing the best conditions, we initiated transesterification of d-glucose with DVA by adding the alkaline protease from Bacillus subtilis (40 mg/mL) as catalyst in a micro-aqueous organic medium (DMF/water: 95/5, v/v) in a shake incubator for 5 days to produce a 6-O-vinyladipoyl d-glucose monomer. The surface properties of this monomer were compared with a commercial product used as an industrial demulsifier. The vinyl sugar ester has then underwent chemical polymerization with potassium persulfate and hydrogen peroxide to produce an amphiphilic polymer with sugar branches. The values obtained for number-average molar mass <Mn>, weigh-average molar mass <Mw> and dispersivity <Mw/Mn> were 1.6 × 10³ g/mol, 2.7 × 10³ g/mol and 1.75, respectively.     

Control on molecular weight reduction of poly(ε-caprolactone) during melt spinning — A way to produce high strength biodegradable fibers

Poly(ε-caprolactone) (PCL) is known for its biocompatibility and biodegradability. These features of PCL have resulted into significant academic as well as industrial research interests for use of this polymer in various areas including biomedical and tissue engineering. Three-dimensional porous scaffolds, controlled drug release systems and nerve guides are some of the forms in which this polymer has been used. Despite these forms, fibers made of PCL have not gained much attention due to PCL's low melting point (57–60 °C) and relatively inferior mechanical properties as compared to poly(L-lactide) (PLA). Also the polymer is sensitive to the process conditions of melt spinning which leads to degradation of PCL when subjected to high temperatures in the presence of air or moisture. Here we present an approach in which addition of a bilactone, bis-(ε-caprolactone-4-yl) (BCY), during melt spinning of PCL resulted into monofilament fibers having tenacity as high as 2500 MPa. The cross-linking of PCL which occurred due to BCY transesterification compensated for molecular weight reduction of the polymer under melt spinning conditions. PCL monofilament fibers thus developed have enhanced thermo-mechanical properties and therefore have high potential to be used in tissue engineering applications in the form of sutures, a mesh or a non-woven.     

Sorption and diffusion of compressed carbon dioxide in polycaprolactone for the development of porous scaffolds

In this work different phenomena related to sorption of carbon dioxide in polycaprolactone (PCL) have been investigated systematically. The use of compressed carbon dioxide is discussed for obtaining porous scaffolds from this biocompatible polymer. In order to determine the plasticization effect of carbon dioxide on the degree of foaming it is necessary to discuss sorption data with respect to morphological features of the polymer at conditions nearby the melting point. The amount of carbon dioxide dissolved and the kinetics of the sorption process are found to depend strongly on temperature and pressure. The solubility takes values of up to 25 wt.% being favoured by a melting and glass transition temperature depression which can be observed along with an enhanced mass transfer rate. In general, CO₂ sorption in PCL increases linearly with pressure. When decompressing, microfoaming occurs which enhances the rate of gas release. Changes in morphology and crystallinity occur as a consequence of the pressure treatment. Compared to the melting temperature at atmospheric pressure there is a dramatic reduction observed under pressure where melting occurs already at a temperature below 40 °C. Even after pressure-treatment, there is a remaining change in melting temperature and crystallinity observed. Optimum conditions for obtaining adequate porous scaffolds of PCL are a relatively slow decompression after treatment at 17 MPa and 35 °C.     

Kinetics of aluminum dross dissolution in sec-butyl alcohol for aluminum sec-butoxide

Al sec-butoxide (ASB) has been used as a precursor for activated aluminas but its cost is higher than any other type of precursor. This study was carried out on the dissolution kinetics for synthesis of the ASB from Al dross waste. The reaction was performed under the molar ratio of Al dross and sec-butyl alcohol (SBA) of 3 mol SBA/mol Al with a catalyst of 10^?3 mol HgI₂/mol Al and three different dissolution temperatures of 80, 90 and 100 °C. The Al reactant was used with Al dross of 3–5 mm size range. As a result of the experiment, the dissolution reaction gave a 65% yield after 24 h. The dissolution mechanism was determined by the shrinking core model assumed by the shape of spherical particles. Especially, the kinetic data were well fitted by a chemical reaction in the model. By the Arrhenius equation, the apparent activation energy was determined to be 40.9 kJ mol^?1 at the given reaction temperatures.     

The effect of point mutations on structure and mechanical properties of collagen-like fibril: A molecular dynamics study

Understanding sequence dependent mechanical and structural properties of collagen fibrils is important for the development of artificial biomaterials for medical and nanotechnological applications. Moreover, point mutations are behind many collagen associated diseases, including Osteogenesis Imperfecta (OI). We conducted a combination of classical and steered atomistic molecular dynamics simulations to examine the effect of point mutations on structure and mechanical properties of short collagen fibrils which include mutations of glycine to alanine, aspartic acid, cysteine, and serine or mutations of hydroxyproline to arginine, asparagine, glutamine, and lysine. We found that all mutations disrupt structure and reduce strength of the collagen fibrils, which may affect the hierarchical packing of the fibrils. The glycine mutations were more detrimental to mechanical strength of the fibrils (WT > Ala > Ser > Cys > Asp) than that of hydroxyproline (WT > Arg > Gln > Asn > Lys). The clinical outcome for glycine mutations agrees well with the trend in reduction of fibril's tensile strength predicted by our simulations. Overall, our results suggest that the reduction in mechanical properties of collagen fibrils may be used to predict the clinical outcome of mutations.     

Preparation and characterization of polyvinyl alcohol hydrogels crosslinked by biodegradable polyurethane for tissue engineering of cartilage

Polyurethane was prepared from hexamethylene diisocyanate (HMDI) and polycaprolactone diol (PCL) with stoichiometry ratio of two in a reactor to form prepolymer. Polyvinyl alcohol (PVA) at PVA/prepolymer ratios of 8, 4, 2 and 1 was crosslinked with the former degradable polyester polyurethane. Fourier transform infrared (FTIR) was employed to confirm polyurethane formation during the course of reactions. FTIR spectrum revealed bands at 1729–1733 cm^? 1 and 3347–3340 cm^? 1 which indicates carbonyl and NH of amine groups, respectively. Polyurethane formation was also confirmed by the absence of the isocyanate peaks (NCO) at 2270 cm^? 1. Dynamic mechanical thermal analysis (DMTA) showed that by increasing prepolymer concentration glass transition temperature decreases from 26 °C for PVA to 19 °C for sample with PVA/prepolymer ratio of 4 and then it rises up to 31 °C. Water uptake measurements illustrated about four fold reduction in swelling ratio of PVA after crosslinking and the sample with equal amounts of PVA and PPU had water uptake of 100%, close to that of a natural cartilage and much less than PVA (425%). All samples had compressive modulus in the range of the articular cartilage (1.9–14.4 MPa). The morphology of the isolated cells on the samples was evaluated by scanning electron microscopy (SEM) and revealed cell attachment and proliferation. The cell viability (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT) and GAG expression (dimethylmethylene blue, DMMB) assays with human chondrocytes on the sample with PVA/prepolymer ratio of one showed about 14 and 33% increase in cell viability and GAG expression after 14 days of culture compare to the PVA, respectively.     

Novel,simple and reproducible method for preparation of composite hierarchal porous structure scaffolds

Demand to develop a simple and adaptable method for preparation the hierarchical porous scaffolds for bone tissue regeneration is ever increasing. This study presents a novel and reproducible method for preparing the scaffolds with pores structure spanning from nano, micro to macro scale. A macroporous Sr-Hardystonite (Sr–Ca₂ZnSi₂O₇, Sr–HT) scaffold with the average pore size of ~ 1200 μm and porosity of ~ 95% was prepared using polymer sponge method. The struts of the scaffold were coated with a viscous paste consisted of salt (NaCl) particles and polycaprolactone (PCL) to provide a layer with thickness of ~ 300–800 μm. A hierarchical porous scaffold was obtained with macro, micro and nanopores in the range of 400–900 μm, 1–120 μm and 40–290 nm, after salt leaching process. These scales could be easily adjusted based on the starting foam physical characteristics, salt particle size, viscosity of the paste and salt/PCL weight ratio.     

Enhanced water-solubility of Licorice extract microparticle prepared by antisolvent precipitation process

In this study, Licorice extract (LE) microparticles were successfully prepared using antisolvent precipitation process. Ethyl acetate and dimethyl sulfoxide, were used as the antisolvent and solvent, respectively. By means of orthogonal experimental design, the influences of several process parameters on the mean particle size (MPS) were investigated. The concentration range of the LE solution, the volume ratio of solvent to antisolvent, dripping speed, and temperature were 4.3–34.5 mg/mL, 1:1–1:12, 1–10 mL/min, and 20–35 °C, respectively. Based on the above orthogonal experiments, the optimum antisolvent precipitation process conditions were found to be: temperature 20 °C, concentration of the LE solution 17.2 mg/mL, volume ratio of solvent to antisolvent 1:4, dripping speed 10 mL/min. The LE microparticles were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform-infrared (FT-IR) spectroscopy, thermal gravimetric analysis (TG), differential scanning calorimetry (DSC), HPLC analysis and dissolution test. And the morphology, crystalline state and chemical structure, drug purity, dissolution rate and bioavailability of LE microparticles were investigated. Under optimum antisolvent precipitation process conditions, the MPS of LE microparticles reached to 85.3 nm, and with uniform distribution. And the LE microparticles had the same chemical structure as the unprocessed drug, but the crystallinity was reduced, purity was increased. Furthermore, the water solubility increased from 4.82 mg/mL to 16.10 mg/mL, and bioavailability is increased by 64.36%.     

Adsorption of collagen to indium oxide nanoparticles and infrared emissivity study thereon

Adsorption of collagen to indium oxide nanoparticles was carried out in water–acetone solution at volumetric ratio of 1:1 with pH value varying from 3.2 to 9.3. As indicated by TGA, maximum collagen adsorption to indium oxide nanoparticles occurred at pH of 3.2. It was proposed that noncovalent interactions such as hydrogen bonding, hydrophilic and electrostatic interactions made main contributions to collagen adsorption. The IR emissivity values (8–14 μm) of collagen-adsorbed indium oxide nanoparticles decreased significantly compared to either pure collagen or indium oxide nanoparticles possibly due to the interfacial interactions between collagen and indium oxide nanoparticles. And the lowest infrared emissivity value of 0.587 was obtained at collagen adsorption of 1.94 g/100 g In₂O₃. On the chance of improved compatibility with organic adhesives, the chemical activity of adsorbed collagen was further confirmed by grafting copolymerization with methyl methacrylate by formation of polymer shell outside, as evidenced by IR spectrum and transmission electron microscopy.     

Template-assisted electrohydrodynamic atomization of polycaprolactone for orthopedic patterning applications

This paper presents the development of the novel deposition of biodegradable polycaprolactone (PCL) polymer patterns on a metallic substrate using a jet spraying technique, template-assisted electrohydrodynamic atomization (TAEA), at ambient temperature. The structure of patterns was controlled by systematically varying the polymer concentration (2–15 wt.%) and the flow rate (1–25 μl min^? 1). Polymer deposition was carried out in the stable cone-jet mode to precisely control the surface structure and morphology. The patterns were studied by optical microscopy, scanning electron microscopy and profilometry, and a high degree of control over the pattern geometry and thickness was achieved by varying the spraying time. The hardness and the effective elastic modulus of the polymer patterns were estimated using nanoindentation. The effect of load, loading rate and the holding time on the hardness and effective elastic modulus was derived. Optimal results were obtained with 5 wt.% PCL in DMAC solution sprayed within the stable cone-jet mode operating window at a flow rate of 15 μl min^? 1 for 300 s at 11.1 kV with a working distance of 60 mm. Hexagonal patterns were well-defined and repeatable with thickness of ~ 34 μm. The hardness is 1.6 MPa at a loading rate of 0.1 μN/s and nearly halved when the load rate was increased to 1 μN/s. The effective elastic modulus of ~ 12 MPa is obtained for a load rate of 0.1 μN/s.     

Electrospun nanofibers composed of poly(ε-caprolactone) and polyethylenimine for tissue engineering applications

Poly(ε-caprolactone) (PCL) electrospun nanofibers have been reported as a scaffold for tissue engineering application. However, high hydrophobicity of PCL limits use of functional scaffold. In this study, PCL/polyethylenimine (PEI) blend electrospun nanofibers were prepared to overcome the limitation of PCL ones because the PEI as a cationic polymer can increase cell adhesion and can improve the electrospinnability of PCL. The structure, mechanical properties and biological activity of the PCL/PEI electrospun nanofibers were studied. The diameters of the PCL/PEI nanofibers ranged from 150.4 ± 33 to 220.4 ± 32 nm. The PCL/PEI nanofibers showed suitable mechanical properties with adequate porosity and increased hydrophilic behavior. The cell adhesion and cell proliferation of PCL nanofibers were increased by blending with PEI due to the hydrophilic properties of PEI.     

Dipyridamole recognition and controlled release by uniformly sized molecularly imprinted nanospheres

We used novel synthetic conditions of precipitation polymerization to obtain uniformly sized molecularly imprinted nanospheres of dipyridamole for application in the design of new drug delivery systems. In addition, the morphology, drug release, and binding properties of molecularly imprinted polymers (MIPs) were studied, and the effects of morphology on other properties were investigated. The MIPs prepared by acetonitrile/chloroform (19:1, v/v) were uniformly sized nanospheres with an average mean diameter of approximately 88 nm at a wetted state, 50 nm at a dry state, and a polydispersity index of 0.062. The imprinted nanospheres showed excellent binding properties and had 62.7% of template binding compared with 17.1% of its blank polymer. The imprinted nanospheres with 67.5 (mg template/of polymer) of binding capacity had better imprinting efficiency than the 50.5% of binding capacity shown by irregularly shaped MIP particles that were prepared by chloroform. The molecular binding abilities of imprinted nanospheres in human serum were evaluated by HPLC analysis (binding about 77% of dipyridamole). Results from release experiments of MIPs showed a very slow, controlled, and satisfactory release of dipyridamole. The loaded drug was released up to 99% in 17 days for nanospheres and 22 days for irregularly shaped particles.     

Detection of growth factor binding to gelatin and heparin using a photonic crystal optical biosensor

Drug–carrier interactions are important to protein controlled release systems to protect the protein from denaturation and ensure properly timed release. A novel photonic crystal biosensor was used to investigate a gelatin–protein controlled release system to determine the amount of protein bound to the carrier at physiological conditions. The Biomolecular Interaction Detection (BIND) system reflects a narrow band of wavelengths when white light is shone incident to the grating. As mass is deposited onto the surface, the peak wavelength value is shifted due to changes in the optical density of the biosensor. The BIND system was used to detect the binding of growth factors onto acidic gelatin, basic gelatin, and heparin on the sensor surface. Through a series of experiments, including functionalizing the sensor, adjusting the ionic strength of the solution, adjusting the substrate concentration, and minimizing non-specific signal, the adsorption of the gelatins and heparin on the sensor was enhanced. The binding interaction of recombinant human transforming growth factor (rhTGF)-β1 and bone morphogenetic protein (rhBMP)-2 with the two types of gelatin and heparin were investigated. The strength of the interaction between rhTGF-β1 and the substrates is in the following order: heparin > acidic gelatin > basic gelatin. RhBMP-2 bound to the substrates but with less intensity than TGF-β1: heparin > basic gelatin > acidic gelatin. This work provides support for the controlled release mechanism through degradation of the gelatin carrier.