Porous CDHA microspheres laden brushite-based injectable bone substitutes for improved bone regeneration.

Porous CDHA microspheres were incorporated into innovative injectable calcium phosphate cement (CPC) to enhance the rate of degradation and bioactivity of bone regeneration. With varying content of CDHA microspheres, the final setting time varied between 12 and 17 min, which is adequate for surgeons to accomplish the implantation. Compressive strength ranged between 6 and 8 MPa, until the addition of porous CDHA microsphere into CPC reached 20 vol %, but decreased dramatically after 30 vol % addition. Therefore, CPC with 20 vol % addition of porous CDHA microspheres was found appropriate for in vitro degradation and cytocompatibility studies. Histological assessment identified new bone formation around the injected bone substitute without significant inflammatory reactions. In vivo analysis of rat femoral defects revealed a threefold higher bone formation in CPC/CDHA 20 vol % than in CPC, due to the more cell migration and penetration into CPC by the existence of porous CDHA microspheres. Based on the promising results obtained, this novel injectable bone substitute may be useful in bone regeneration.

Development of a novel polycaprolactone based composite membrane for periodontal regeneration using spin coating technique.

Guided bone regeneration (GBR) is known to prevent the development of soft tissue on the defect sites as well as support the new bone formation on the other end. In the present study, we developed a multilayer biodegradable membrane for GBR applications. The multilayer membrane is primarily composed of ß-tricalcium phosphate (TCP), polycaprolactone (PCL), and hyaluronic acid (HA), prepared by the spin-coating method. The triple layer system has PCL-TCP composite layer on top, a PCL layer in the middle, and PCL-HA as the bottom layer. The characterization of the PCL-TCP/PCL/PCL-HA by various techniques such as SEM, EDS, XRD, and FT-IR supported the uniform formation of the triple layers with an overall thickness of ∼ 72 µm. Multilayer composite membrane showed excellent physical parameters; neutral pH, high hydrophilicity, high swelling rate, low degradation rate, and high apatite formation after immersion in simulated body fluid (SBF) for 14 days. The multilayer membrane also exhibited biocompatibility which is evident by MTT assay and confocal images. The results suggested that the multilayer composite membrane has the potential for GBR applications.

Comparative study on biodegradation and biocompatibility of multichannel calcium phosphate based bone substitutes.

The objective of this study was to fabricate multichannel biphasic calcium phosphate (BCP) and ß-tricalcium phosphate (TCP) bone substitutes and compare their long-term biodegradation and bone regeneration potentials. Multi-channel BCP and TCP scaffolds were fabricated by multi-pass extrusion process. Both scaffolds were cylindrical with a diameter of 1-mm, a length of 1-mm, and seven interconnected channels. Morphology, chemical composition, phase, porosity, compressive strength, ion release behavior, and in-vitro biocompatibility of both scaffolds were studied. In-vivo biodegradation and bone regeneration efficacies of BCP and TCP were also evaluated using a rabbit model for 1 week, 1 month, and 6 months. BCP exhibited superior compressive strength compared to TCP scaffold. TCP showed higher release of both calcium ions and phosphorous ions than BCP in SBF solution. Both scaffolds showed excellent in-vitro biocompatibility and upregulated the expression of osteogenic markers of MC3T3-E1 cells. In-vivo studies revealed that both cylindrical TCP and BCP scaffolds were osteoconductive and supported new bone formation. Micro-CT data showed that the bone-regeneration efficacy of TCP was higher at one month and at six months after implantation. Histological examination confirmed that TCP degraded faster and had better bone regeneration than BCP after 6 months.

Bone regeneration of multichannel biphasic calcium phosphate granules supplemented with hyaluronic acid.

The present work is to investigate the efficiency of hyaluronic acid (HyA) supplemented biphasic calcium phosphate (BCP) injectable granule to promote the bone regeneration. The effect of adding HyA to the multichannel BCP granule (MCG-HyA) was studied in terms of morphology, chemical structure, porosity, in-vitro and in-vivo biocompatibility, RT-PCR, western blot and compared with MCG. The addition of HyA to MCG successfully made the granules injectable type. In-vivo studies in rabbit model showed an enhancement in bone formation after 4 weeks of implantation and better handling characteristics for MCG-HyA than MCG. RT-PCR and Western Blotting studies revealed that MCG-HyA significantly unregulated the osteogenic gene and protein expressions respectively. Our results indicated that MCG-HyA could be used as a promising injectable bone substitute in clinical applications.

In vitro and in vivo assessment of biomedical Mg-Ca alloys for bone implant applications.

BACKGROUND: Magnesium (Mg)-based alloys are considered to be promising materials for implant application due to their excellent biocompatibility, biodegradability, and mechanical properties close to bone. However, low corrosion resistance and fast degradation are limiting their application. Mg-Ca alloys have huge potential owing to a similar density to bone, good corrosion resistance, and as Mg is essential for Ca incorporation into bone. The objective of the present work is to determine the in vitro degradation and in vivo performance of binary Mg- xCa alloy ( x = 0.5 or 5.0 wt%) to assess its usability for degradable implant applications. METHODS: Microstructural evolutions for Mg- xCa alloys were characterized by optical, SEM, EDX, and XRD. In vitro degradation tests were conducted via immersion test in phosphate buffer saline solution. In vivo performance in terms of interface, biocompatibility, and biodegradability of Mg- xCa alloys was examined by implanting samples into rabbit femoral condyle for 2 and 4 weeks. RESULTS: Microstructural results showed the enhancement in intermetallic Mg2Ca phase with increase in Ca content. Immersion tests revealed that the dissolution rate varies linearly, with Ca content exhibiting more hydrogen gas evolution, increased pH, and higher degradation for Mg-5.0Ca alloy. In vivo studies showed good biocompatibility with enhanced bone formation for Mg-0.5Ca after 4 weeks of implantation compared with Mg-5.0Ca alloy. Higher initial corrosion rate with prolonged inflammation and rapid degradation was noticed in Mg-5.0Ca compared with Mg-0.5Ca alloy. CONCLUSIONS: The results suggest that Mg-0.5Ca alloy could be used as a temporary biodegradable implant material for clinical applications owing to its controlled in vivo degradation, reduced inflammation, and high bone-formation capability.

Development and properties of duplex MgF2/PCL coatings on biodegradable magnesium alloy for biomedical applications.

The present work addresses the performance of polycaprolactone (PCL) coating on fluoride treated (MgF2) biodegradable ZK60 magnesium alloy (Mg) for biomedical application. MgF2 conversion layer was first produced by immersing Mg alloy substrate in hydrofluoric acid solution. The outer PCL coating was then prepared using dip coating technique. Morphology, elements profile, phase structure, roughness, mechanical properties, invitro corrosion, and biocompatibility of duplex MgF2/PCL coating were then characterized and compared to those of fluoride coated and uncoated Mg samples. The invivo degradation behavior and biocompatibility of duplex MgF2/PCL coating with respect to ZK60 Mg alloy were also studied using rabbit model for 2 weeks. SEM and TEM analysis showed that the duplex coating was uniform and comprised of porous PCL film (~3.3 µm) as upper layer with compact MgF2 (~2.2 µm) as inner layer. No significant change in microhardness was found on duplex coating compared with uncoated ZK60 Mg alloy. The duplex coating showed improved invitro corrosion resistance than single layered MgF2 or uncoated alloy samples. The duplex coating also resulted in better cell viability, cell adhesion, and cell proliferation compared to fluoride coated or uncoated alloy. Preliminary invivo studies indicated that duplex MgF2/PCL coating reduced the degradation rate of ZK60 Mg alloy and exhibited good biocompatibility. These results suggested that duplex MgF2/PCL coating on magnesium alloy might have great potential for orthopedic applications.

A novel hybrid multichannel biphasic calcium phosphate granule-based composite scaffold for cartilage tissue regeneration.

The objective of the present study was to develop a novel hybrid multichannel biphasic calcium phosphate granule (MCG)-based composite system for cartilage regeneration. First, hyaluronic acid-gelatin (HG) hydrogel was coated onto MCG matrix (MCG-HG). Poly(lactic-co-glycolic acid) (PLGA) microspheres was separately prepared and modified with polydopamine subsequent to BMP-7 loading (B). The surface-modified microspheres were finally embedded into MCG-HG scaffold to develop the novel hybrid (MCG-HG-PLGA-PD-B) composite system. The newly developed MCG-HG-PLGA-PD-B composite was then subjected to scanning electron microscopy, energy dispersive X-ray spectroscopy, Fourier Transform infrared spectroscopy, porosity, compressive strength, swelling, BMP-7 release and in-vitro biocompatibility studies. Results showed that 60% of BMP-7 retained on the granular surface after 28 days. A hybrid MCG-HG-PLGA-PD-B composite scaffold exhibited higher swelling and compressive strength compared to MCG-HG or MCG. In-vitro studies showed that MCG-HG-PLGA-PD-B had improved cell viability and cell proliferation for both MC3T3-E1 pre-osteoblasts and ATDC5 pre-chondrocytes cell line with respect to MCG-HG or MCG scaffold. Our results suggest that a hybrid MCG-HG-PLGA-PD-B composite scaffold can be a promising candidate for cartilage regeneration applications.

Comparative Bone Regeneration Potential Studies of Collagen, Heparin, and Polydopamine-Coated Multichannelled BCP Granules.

The current study is a comparative assessment of the bone regeneration potentiality of bone substitutes composed of bioactive polymer-coated biphasic calcium phosphate (BCP) granules. The bone substitutes were primarily composed of multichanneled BCP granules, which were coated separately with biochemical polymer coatings, namely collagen, heparin, or polydopamine (PD), using chemical methods. The morphologic features and chemical structure of the granules and surface coatings were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy, respectively. The biological characterizations in terms of cellular interaction with the bone substitutes in vitro (MTT assay and proliferation) and in vivo (bone regeneration efficacy) were performed and compared with uncoated BCP granules. It was found that PD-coated BCP granules were superior to the others with respect to promoting more rapid healing. Therefore, PD multichannel BCP granule system can be used as a promising injectable bone substitute for clinical applications.

Incorporation of BMP-2 loaded collagen conjugated BCP granules in calcium phosphate cement based injectable bone substitutes for improved bone regeneration.

The objective of the present study was to incorporate surface modified porous multichannel BCP granule into CPC to enhance its in vivo biodegradation and bone tissue growth. The multichannel BCP granule (15wt%) was first coated with collagen subsequent to BMP-2 loading (ccMCG-B). It was then embedded into CPC to form CPC-ccMCG-B system. The newly developed CPC-ccMCG-B system was then examined for SEM, EDX, XRD, setting time, compressive strength, injectability, pH change, BMP-2 release, in vitro as well as in vivo studies and further compared with CPC. Optimized CPC (0.45mL/g) was found based on setting time and compressive strength studies. In vivo studies exhibited improved new bone formation and better degradation of CPC after 2 and 4weeks of implantation as compared to CPC as resulted from effective BMP-2 signaling. Our results suggest that CPC-ccMCG-B system might be used as a promising injectable bone substitutes in clinical applications.