Coral Scaffold Research Articles

Sustainably cultured coral scaffold supports human bone marrow mesenchymal stromal cell osteogenesis

The current gold standard grafting material is autologous bone due to its osteoinductive and osteoconductive properties. Autograft harvesting results in donors site morbidity. Coral scaffolds offer a natural autograft alternative, sharing the density and porosity of human bone. This study investigated the biocompatibility and osteogenic potential of a novel, sustainably grown Pocillopora scaffold with human bone marrow-derived mesenchymal stromal cells (MSCs). The coral-derived scaffold displays a highly textured topography, with concavities of uniform size and a high calcium carbonate content. Large scaffold samples exhibit compressive and diametral tensile strengths in the range of trabecular bone, with strengths likely increasing for smaller particulate samples. Following the in vitro seeding of MSCs adjacent to the scaffold, the MSCs remained viable, continued proliferating and metabolising, demonstrating biocompatibility. The seeded MSCs densely covered the coral scaffold with organized, aligned cultures with a fibroblastic morphology. In vivo coral scaffolds with MSCs supported earlier bone and blood vessel formation as compared to control constructs containing TCP-HA and MSCs. This work characterized a novel, sustainably grown coral scaffold that was biocompatible with MSCs and supports their in vivo osteogenic differentiation, advancing the current repertoire of biomaterials for bone grafting.

Podium Presentation Title: Lesion Features Have No Effect on Outcomes when Comparing a Coral Scaffold vs. Debridement/Micro-Fixation

Podium Presentation Title: Lesion Features Have No Effect on Outcomes when Comparing a Coral Scaffold vs. Debridement/Micro-Fixation

Study on porous coral scaffolds containing a hydroxyapatite layer doped with selenium and their properties.

The repair of bone defects caused by osteosarcoma is still a significant clinical issue, and new scaffolds need to be developed to solve this problem. The ocean is a treasure trove for developing new biomedical materials, and coral is widely thought to be suitable as a scaffold for bone implant materials due to its porous structure and mechanical properties. Selenium is known for its antioxidant and antitumor effects, inducing tumor cell cycle arrest. In this study, we hydrothermally transformed corals to grow a hydroxyapatite layer on the scaffold surface (CHAp) and combined it with selenium to obtain selenium-doped scaffolds (Se-CHAp) without affecting the porous structure of the coral. The research successfully validates their biocompatibility and the antitumor efficacy against 143B osteosarcoma cells. The results indicate that the Se-CHAp scaffolds yielded an obvious inhibitory effect on the proliferation of osteosarcoma cells, highlighting that they have huge prospects for application in biomedical technology.

Hydrothermally treated coral scaffold promotes proliferation of mesenchymal stem cells and enhances segmental bone defect healing.

Introduction: Synthetic hydroxyapatite (HAp) scaffolds have shown promising therapeutic outcomes in both animals and patients. In this study, we aim to evaluate the chemical and physical phenotype, biocompatibility, and bone repair effects of hydrothermally treated coral with natural coral and synthetic HAp. Methods: The phase composition, surface pattern, 3D structures, and porosity of the scaffolds were characterized, and cell viability, proliferation, and osteogenic differentiation of mesenchymal stem cells (MSCs) after seeding onto the scaffold were determined. The scaffolds were implanted into rats to assess their bone repair effects using micro-CT analysis, mechanical testing, and histological staining. Results: The results showed that the phase composition, porous structure, and porosity of hydrothermally treated coral were comparable to pure HAp scaffold. While only the natural coral happens to be dominantly calcium carbonate. Higher cell proliferation and osteogenic differentiation potential were observed in the hydrothermally treated coral scaffold compared to natural coral and pure HAp. Histological results also showed increased new bone formation in the hydrothermally treated coral group. Discussion: Overall, our study suggests that hydrothermal modification enhances the cytocompatibility and therapeutic capacity of coral without altering its physical properties, showing superior effectiveness in bone repair to synthetic HAp.

Printed coral scaffold could help regrow reefs

Printed coral scaffold could help regrow reefs

Effect of Incorporation Platelet Rich Plasma into Synthetic Coral Scaffold toward Epithelial Thickness of Wound Healing

Tissue re-epithelialization is a process in wound healing. The healing process of wounds needs to pass through a phase of inflammatory, proliferation, and remodeling. Tissue engineering is the technology service in the field of medicine that can be applied in wound healing. A scaffold is a place for the molecule signal and cell growth to form new tissue. This study aims to investigate the ability of synthetic coral scaffold incorporated PRP in wound healing. Rattus norvegicus were used and divided into 4 groups, synthetic coral scaffold incorporated PRP, scaffold only, Spongostan, and control. The wound healing was observed on 3, 7, and 14 days by measuring the epithelial thickness. The average epithelium thickness on day 3 has not formed. The highest epithelial thickness on days 7 and 14 are obtained from the synthetic coral scaffolds incorporated by PRP. The result of this study presented that Synthetic coral scaffold incorporated by PRP has the highest ability for wound healing and increases the process of tissue regeneration.

Engineering of axially vascularized bone tissue using natural coral scaffold and osteogenic bone marrow mesenchymal stem cell sheets

IntroductionBlood supply remains one of the obstacles to large bone tissue engineering. This study aimed to generate vascularized bone tissue by inducing axial vascularization into a construct combining natural coral scaffold and a bone marrow mesenchymal stem cells (BMSCs) sheet. Material and methodsIsolated BMSCs were cultured to form an osteogenic cell sheet using a continuous culture method. Natural coral scaffolds were prepared into customized shape with a cylinder of 20 mm length, 8 mm in outer diameter and 5 mm in inner diameter. Then, the freed superficial inferior epigastric vessel of rabbits was first wrapped with a cell sheet, and then inserted into the central passage of the scaffold, after being wrapped with another cell sheet, the complexes were implanted subcutaneously into a rabbit groin area. In contrast, the sheet-scaffold construct that implanted into groin subcutaneous area of the other side of the same rabbit with the distal end of the blood vessel was ligated, which was considered as control. New bone and vascularization formation were evaluated at 12 weeks postoperatively. ResultsThe volume of new bone formation and amount of capillary infiltration in the vascular circulation group were significantly greater than that in the vascular ligation group, which suggested that insertion of axial vessels could significantly promote angiogenesis and osteogenesis of the tissue-engineered bone. ConclusionsThese findings indicate that inserting an arteriovenous bundle into the constructs of mesenchymal stem cell sheet and coral has great potential for clinical applications to repair large bone defects.

The Biological Effects of Coral with Adipose Derived Stem Cells on Mandibular Bony Defect in Albino Rats.

Objective: To evaluate the biological effects of coral scaffold with or without adipose derived stem cells (ADSCs) on healing of bony defects in albino rats. Methods: Fifty four male albino rats weighing approximately 250-300 gm were selected and randomly divided into 3 groups. Each rat received one bone defect on the left side of mandible near the diastema. The defects were either left empty (group A) n=18, treated with coral only (group B) n=18 or received a combination of coral and adipose derived stem cells (group C) n=18 which were harvested from the rats peri renal region. Bone healing was evaluated using heamatoxylin and eosin and trichrome stain, then the results were subjected to digital image analysis followed by one- way ANOVA statistical analysis. Results: Bony defects received the combination of coral scaffold and stem cells show better healing in respect to the quantity and the quality of the newly formed bone. Conclusion: ADSCs loaded onto coral granule scaffold have ameliorating effect on the repair of the mandibular defects in rat model.

Construction of tissue-engineered bone with differentiated osteoblasts from adipose-derived stem cell and coral scaffolds at an ectopic site

Cell sheets from bone marrow mesenchymal stem cells (BMSC) have been widely used in the field of bone tissue engineering, although their source remains a challenging issue. In this study, adipose-derived stem cells (ADSC) were induced to differentiate into osteoblasts, and the incorporation of coral scaffolds with ADSC sheets for bone formation at an ectopic site was also investigated. First, ADSC isolated from inguinal adipose tissue of New Zealand rabbits were cultured for two weeks without passaging under osteogenic induction, and the microstructures of cell sheets were analysed by histological and scanning electron microscope (EM) observation. In addition, the activity of alkaline phosphatase (ALP) and alizarin red staining was also measured to detect their osteogenic ability. Subsequently, ADSC were proved to be able to proliferate well when seeded on the coral scaffolds. Next, coral scaffolds were wrapped in cell sheets to prepare sheet-coral complexes, which were implanted into subcutaneous pockets in nude mice. At eight weeks after implantation, gross examination, microcomputed tomography (MicroCT), and histological analysis were investigated to assess new bone formation. MicroCT scanning and histological analysis showed that there was more highly dense tissue formed in the complex group than control group (p=0.0004). These results indicated that osteoblastic ADSC sheets could be used to construct engineered bone and the incorporation of coral scaffolds with ADSC sheets significantly improved bone formation, providing a newly approach for bone tissue engineering.

Degradation, swelling profile, and gel fraction of synthetic coral scaffold incorporated PRP or PRF

Platelet rich plasma (PRP) and platelet rich fibrin (PRF) are sourced from human blood and have role in bone recovery. Both of them could be incorporated into scaffold for bone regeneration. Synthetic coral scaffold is a natural mimicking of sea coral designed for bone regeneration. It should have bio degradability and bio absorbability that influence new bone formation process. The purpose of this study is to investigate the degradation, swelling profile and gel fraction of synthetic coral scaffold incorporated with PRP or PRF. Synthetic coral scaffold consists of gelatin, calcium carbonate, and sodium nitrate as dispersant agent. It is divided into 3 groups, incorporated PRP, PRF, and no-incorporation. Scaffold was soaked in phosphate buffer saline (PBS) and incubated in 37° C for 24 hours. Scaffold weight was measured in every 30 minutes to observe the swelling profile and gel fraction. Degradation profile was observed after 1, 3, 6, 24, 48, 72, and 96 hours of soaking. Acceleration of degradation was measured after soaking with HC1 1N for 1, 3, 6, 24, 48, 72, 96 hours until the scaffold ran out. The result shows significant differences among PRP, PRF, and control group. PRP incorporation has slow degradation, good swelling and higher gel fraction that presents the strengthening of scaffold structure.

The Porosity and Human Gingival Cells Attachment of Synthetic Coral Scaffold for Bone Regeneration

Porosity and interconnectivity play an important role in the success of tissue engineering because it affects cells to live and grow. Coral has been used as a bone replacement material because the structures resemble bone and have mechanical bone properties. In this study, the synthetic coral scaffold was developed to mimic the natural coral. This study aims to investigate the porosity of the scaffold and its biocompatibility while it is attached to human gingival cells. Synthetic coral scaffold in various compositions were prepared, porosity percentage measurement and human gingival cell attachment test were done. An optimum ratio of the scaffold with gelatin: CaCO3, having the highest porosity and cell attachment is obtained in 5:5. The result of this study presented that synthetic coral scaffold could provide the microenvironment to cells for life because it is supported by the highest percentage of porosity.

Characteristic of Synthetic Coral Scaffold for Cell Environment

Introduction. Synthetic coral scaffold is fabricated the mimicking of natural sea coral as a scaffold for bone regeneration [1]. Scaffold is performing functions as a micro environment for cells attachment, growth, proliferates, differentiates until it can form new bone tissue. The proper design is needed to produce the scaffold [2]. The purpose of this study was to investigate the characteristics of synthetic coral scaffold for micro environment of cells by observing cell attachment, hydrophobicity, and scaffold porosity. Experimental. Synthetic coral scaffold consists of bovine gelatin dan CaCO3 by weight, the concentration that be used are 4:6. Sodium citrate is used as dispersant. Thick film like scaffold was prepared for this study [1,3]. Vero cell line was used for observing cell attachment to investigate the biocompatibility the scaffold. The hydrophobicity was observed with distilled water droplets dripped on the scaffold surface, be analyzed in a photograph taken by the camera and then measured the angle. Percentage of porosity was measured using Archimedes law in absolute ethanol. Results and Discussion. Vero cells attached successfully into scaffold. Cell viability percentage is 91,77 % from the absorbance value of the MTT assay. It presented that the scaffold has biocompatibility character. However, the percentage of porosity is 55,85%, so the scaffold has enough porosity for cell attachment. Porosity serves for the diffusion of nutrients, gases and removes the residual metabolism resulting from cell activity that has grown on scaffold. The good porosity value of the scaffold is 50-90%. The higher the porosity value the better the scaffold. Hydrophobicity scaffold appears from the contact angle of 81.4°, the cohesion is greater than the adhesion. This shows the greater synthetic coral scaffold hydrophobicity, which is affected by surface roughness from scaffold porosity. The greater hydrophobicity will also prolong the degradation of the scaffold, thereby enabling cells to proliferate, differentiate and produce bone matrix. Conclusions. Synthetic coral scaffold provides the micro environment for cell, high hydrophobicity allows longer degradation for proliferation and differentiation of bone cells, and porosity that allows cells to be inserted within the scaffold.

Diversity of the sponge fauna associated with white coral banks from two Sardinian canyons (Mediterranean Sea)

AbstractThe three-dimensional coral scaffolds formed by the skeletons of the cold-water corals Madrepora oculata and Lophelia pertusa represent an important deep-sea hard substratum and create an optimal shelter for a rich associated fauna in which the contribution of Porifera has still not been fully considered. The taxonomic analysis of sponges collected from two Sardinian canyons (Nora and Coda Cavallo, 256–408 m) and associated with the dead coral matrix resulted in 28 species, including new records for the Mediterranean Sea, Italian fauna or Central Tyrrhenian Sea. In addition, for many species this is the first finding associated with the coral framework or the first documentation of the in situ morphology. The taxonomic comparison with sponge assemblages associated with coral frameworks from Santa Maria di Leuca, Strait of Sicily and Bari Canyon, gave the opportunity to evaluate the similarities among geographically separated banks. Overall, the percentage of exclusive species (recorded only in one site), is very high (81%) and only one species is shared by all four sites, suggesting a low connectivity among the sponge communities. The percentage of shared species is higher for the Maltese community, supporting the role of the Sicily Channel as a crossroads between the communities of the eastern and western Mediterranean basins. Here, 55% of the sponges associated to the coral framework are also reported in shallow-water coralligenous assemblages, indicating a high bathymetric connectivity as well as an ecological plasticity allowing these species to occupy a wide range of small, dark refuges.

Periodontal Regeneration Using Cultured Coral Scaffolds in Class II Furcation Defects in Dogs

Periodontal Regeneration Using Cultured Coral Scaffolds in Class II Furcation Defects in Dogs

Degradation profile of synthetic coral scaffold in cell culture media

<p>The scaffold is one of the factors in tissue engineering that determine the success of bone regeneration. The important characteristic of the scaffold is able to degrade gradually. In vitro study using cells, the scaffold will be exposed to culture media. Therefore, degradation profile for scaffold needs to be examined. This study aims to investigate the degradation profile of synthetic coral scaffold in cell culture media using pH measurement. The method used the synthetic coral scaffolds were prepared from denaturalized collagen (gelatin) and calcium carbonate (calcite) with a concentration of 5:5 and 4:6 weight % in aqua dest. The scaffold<span lang="IN" style="">s</span> were fabricated in membrane thick film which was then physically crosslinked.  <span lang="IN" style="">The </span>10 % <span lang="IN" style="">of gelatin scaffold was used as a control.</span>  The scaffolds were incubated in cell culture media (non-phenol red Dulbecco’s Modified Eagle Medium) for 1 until 8 days, and pH changes of the medium were measured. As the result, Profile of degradation on day 1 to day 4 showed the 5:5 scaffold had the smallest degradation. The results indicated the significant different between scaffold concentration in day 1 (p=0.005), 5th(p=0.03), and 6th day (p=0.011). At the end of incubated days, the pH changed but not significantly different. LSD showed the significant differences between scaffold (5:5 and 4:6) with control and no significant difference between 2 concentrations of the scaffold. The conclusion of this study is the synthetic coral scaffold degraded gradually until the end incubation time and between concentration had different degradation profile in the early incubation time using pH measurement.<br></p>

Low‐dose BMP‐2 and MSC dual delivery onto coral scaffold for critical‐size bone defect regeneration in sheep

Tissue-engineered constructs (TECs) combining resorbable calcium-based scaffolds and mesenchymal stem cells (MSCs) have the capability to regenerate large bone defects. Inconsistent results have, however, been observed, with a lack of osteoinductivity as a possible cause of failure. This study aimed to evaluate the impact of the addition of low-dose bone morphogenetic protein-2 (BMP-2) to MSC-coral-TECs on the healing of clinically relevant segmental bone defects in sheep. Coral granules were either seeded with autologous MSCs (bone marrow-derived) or loaded with BMP-2. A 25-mm-long metatarsal bone defect was created and stabilized with a plate in 18 sheep. Defects were filled with one of the following TECs: (i) BMP (n = 5); (ii) MSC (n = 7); or (iii) MSC-BMP (n = 6). Radiographic follow-up was performed until animal sacrifice at 4 months. Bone formation and scaffold resorption were assessed by micro-CT and histological analysis. Bone union with nearly complete scaffold resorption was observed in 1/5, 2/7, and 3/6 animals, when BMP-, MSC-, and MSC-BMP-TECs were implanted, respectively. The amount of newly formed bone was not statistically different between groups: 1074 mm3 [970-2478 mm3 ], 1155 mm3 [970-2595 mm3 ], and 2343 mm3 [931-3276 mm3 ] for BMP-, MSC-, and MSC-BMP-TECs, respectively. Increased scaffold resorption rate using BMP-TECs was the only potential side effect observed. In conclusion, although the dual delivery of MSCs and BMP-2 onto a coral scaffold further increased bone formation and bone union when compared to single treatment, results were non-significant. Only 50% of the defects healed, demonstrating the need for further refinement of this strategy before clinical use. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2637-2645, 2017.

Hydrothermally converted marine coral scaffold promotes segmental bone defect healing in rats

Hydrothermally converted marine coral scaffold promotes segmental bone defect healing in rats

A Selective Cell Population from Dermis Strengthens Bone Regeneration.

Finding appropriate seed cells for bone tissue engineering remains a significant challenge. Considering that skin is the largest organ, we hypothesized that human bone morphogenetic protein receptor type IB (BmprIB)+ dermal cells could have enhanced osteogenic capacity in the healing of critical‐sized calvarial defects in an immunodeficient mouse model. In this study, immunohistochemical staining revealed that BmprIB was expressed throughout reticular dermal cells; the positive expression rate of BmprIB was 3.5% ± 0.4% in freshly separated dermal cells, by flow cytometry. Furthermore, in vitro osteogenic capacity of BmprIB+ cells was confirmed by osteogenic‐related staining and marker gene expression compared with unsorted dermal cells. In vivo osteogenic capacity was demonstrated by implantation of human BmprIB+ cell/coral constructs in the treatment of 4‐mm diameter calvarial defects in an immunodeficient mouse model compared with implantation of unsorted cell/coral constructs and coral scaffold alone. These results indicate that the selective cell population BmprIB from human dermis is a promising osteogenic progenitor cell that can be a large‐quantity and high‐quality cell source for bone tissue engineering and regeneration. Stem Cells Translational Medicine 2017;6:306–315

Human Mesenchymal Stem Cells Behavior on Synthetic Coral Scaffold

For bone tissue engineering, corals have long history to be used as scaffold to promote bone regeneration. However the use of a lot of corals may damage their habitates. For this reason, a strategy to mimic coral in a synthetic form is needed. As an ideal scaffold, synthetic coral must provide structure and initial support for cell attachment and proliferation. The aim of this study was to investigate the attachment and proliferation of human Mesenchymal Stem Cells (h-MSC) on synthetic coral scaffold, to provide information on the behavior of h-MSC on the designated scaffold. Synthetic coral scaffolds were prepared from bovine gelatine and CaCO3 with 5:5 in 10% w/v concentration in aquadest. Sodium citrate was used as dispersant in the suspension. Gelatin-CaCO3 suspension was moulded in a plastic cover of 24 well plate, then freezed at -20°C for 24 hours, freeze dried for 24 hours and continued by dehydrothermal crosslinking for 72 hours. After the fabrication, synthetic coral scaffolds were subjects to cover the bottom of the well for cell culture. Human Mesenchymal Stem Cells (h-MSC) were seeded and divided into 2 groups, control group without scaffold and the one with scaffold. All groups were incubated for 3, 6, and 24 hours. Cells attatchment were determined by deduction of the cells unattached from total cells seeding. Proliferation of h-MSC were done in 3 groups ie., control group without scaffold, scaffold only and scaffold incorporated Platelet Rich Plasma (PRP) in the bottom of well. All groups were incubated for 24, 48 and 72 hours. Human Mesenchymal Stem Cells attached faster to synthetic coral scaffold than the control. Its proliferation behavior was faster in the scaffold incorporated PRP, showing better interaction of scaffold and cells with the incorporation of morphogenetic factor.

Comparative study of the osteogenic ability of four different ceramic constructs in an ectopic large animal model

Tissue-engineered constructs combining bone marrow mesenchymal stem cells with biodegradable osteoconductive scaffolds are very promising for repairing large segmental bone defects. Synchronizing and controlling the balance between scaffold-material resorption and new bone tissue formation are crucial aspects for the success of bone tissue engineering. The purpose of the present study was to determine, and compare, the osteogenic potential of ceramic scaffolds with different resorbability. Four clinically relevant granular biomaterial scaffolds (specifically, Porites coral, Acropora coral, beta-tricalcium phosphate and banked bone) with or without autologous bone marrow stromal cells were implanted in the ectopic, subcutaneous-pouch sheep model. Scaffold material resorption and new bone formation were assessed eight weeks after implantation. New bone formation was only detected when the biomaterial constructs tested contained MSCs. New bone formation was higher in the Porites coral and Acropora coral than in either the beta-tricalcium phosphate or the banked bone constructs; furthermore, there was a direct correlation between scaffold resorption and bone formation. The results of the present study provide evidence that, among the biomaterials tested, coral scaffolds containing MSCs promoted the best new bone formation in the present study.