Nickel-ion substituted hydroxyapatite matrices for metal-affinity chromatographic purification of recombinant proteins.

Hydroxyapatite (HAp) is a calcium phosphate ceramic, widely used as a matrix for protein chromatography. The crystal structure of HAp is amenable to a wide range of substitutions, thus allowing for the alteration of its properties. In this study, nickel-ion substituted HAp (NiSHAp) was synthesized using a wet-precipitation method, followed by spray drying. This resulted in the structural incorporation of nickel ions within well-defined microspheres, which were suitable for chromatographic applications. The chromatographic experiments were conducted with NiSHAp and compared with spray-dried hydroxyapatite (SHAp) matrices. Protein purification experiments were conducted using refolded recombinant L-asparaginase (L-Asp), which was produced as inclusion bodies in Escherichia coli. The results showed that NiSHAp effectively adsorbed L-Asp, which was selectively eluted using a phosphate buffer, surpassing the efficiency of imidazole-based elution. In contrast, SHAp showed weaker binding and lower selectivity. The significance of this study lies in developing a scalable NiSHAp matrix for protein purification, especially for large-scale applications. The NiSHAp matrix offers a cost-effective alternative to commercial immobilized metal affinity chromatography (IMAC) adsorbents, especially for purifying His-tagged proteins. This innovative approach exhibits the advantages of mixed-mode chromatography by combining the properties of hydroxyapatite and IMAC in a single matrix, with the potential of improved industrial-scale protein purification.

Functionally multifaceted alginate/curdlan/agarose-based bilayer fibro-porous dressings for addressing full-thickness diabetic wounds.

Full-thickness diabetic wounds are chronic injuries characterized by bleeding, excessive exude, and prolonged inflammation. Single-layer dressings fail to address their disturbed pathophysiology. Therefore, bilayer dressings with structural and compositional differences in each layer have gained attention. We hypothesized that natural polymer (alginate, curdlan, and agarose) based bilayer dressings with inherent healing properties could effectively resolve these issues. Hence, bilayer dressings were fabricated by electrospinning curdlan/agarose/ polyvinyl alcohol blend (top layer) on an alginate/agarose/polyvinyl alcohol-based lyophilized porous (bottom) layer. Ciprofloxacin was incorporated in both layers as a potential antibacterial drug. The bilayer dressing exhibited high swelling (~1300 %), biocompatibility (>90 % with NIH 3T3 and L929 mouse fibroblasts), and hemocompatibility (hemolysis <5 %). In vitro, scratch assay revealed a faster wound closure (~ 95-100 %) than control. Inhibition zone assay revealed antibacterial activity against Staphylococcus aureus and Escherichia coli. Real-time (in vitro) gene expression experiments performed using human THP-1 macrophages exhibited a significant increase in anti-inflammatory cytokines (4.51 fold in IL-10) and a decrease in pro-inflammatory cytokines (1.42 fold in IL-6) in comparison to lipopolysaccharide. Thus, fabricated dressings with high swelling, hemostatic, immunomodulatory, and antibacterial characteristics can serve as potential multifunctional and sustainable templates for healing full-thickness diabetic wounds.

Perspectives of nanofibrous wound dressings based on glucans and galactans - A review.

Wound healing is a complex and dynamic process that needs an appropriate environment to overcome infection and inflammation to progress well. Wounds lead to morbidity, mortality, and a significant economic burden, often due to the non-availability of suitable treatments. Hence, this field has lured the attention of researchers and pharmaceutical industries for decades. As a result, the global wound care market is expected to be 27.8 billion USD by 2026 from 19.3 billion USD in 2021, at a compound annual growth rate (CAGR) of 7.6 %. Wound dressings have emerged as an effective treatment to maintain moisture, protect from pathogens, and impede wound healing. However, synthetic polymer-based dressings fail to comprehensively address optimal and quick regeneration requirements. Natural polymers like glucan and galactan-based carbohydrate dressings have received much attention due to their inherent biocompatibility, biodegradability, inexpensiveness, and natural abundance. Also, nanofibrous mesh supports better proliferation and migration of fibroblasts because of their large surface area and similarity to the extracellular matrix (ECM). Thus, nanostructured dressings derived from glucans and galactans (i.e., chitosan, agar/agarose, pullulan, curdlan, carrageenan, etc.) can overcome the limitations associated with traditional wound dressings. However, they require further development pertaining to the wireless determination of wound bed status and its clinical assessment. The present review intends to provide insight into such carbohydrate-based nanofibrous dressings and their prospects, along with some clinical case studies.

Fabrication and evaluation of agarose-curdlan blend derived multifunctional nanofibrous mats for diabetic wounds.

Diabetic wounds with complex pathophysiology significantly burden the wound care industry and require novel management strategies. In the present study, we hypothesized that agarose-curdlan based nanofibrous dressings could be an effective biomaterial for addressing diabetic wounds due to their inherent healing properties. Hence, agarose/curdlan/polyvinyl alcohol based nanofibrous mats loaded with ciprofloxacin (0, 1, 3, and 5 wt%) were fabricated using an electrospinning technique with water and formic acid. In vitro evaluation revealed the average diameter of the fabricated nanofibers between 115 and 146 nm with high swelling (~450-500 %) properties. They exhibited enhanced mechanical strength (7.46 ± 0.80 MPa -7.79 ± 0.007 MPa) and significant biocompatibility (~90-98 %) with L929 and NIH 3T3 mouse fibroblasts. In vitro scratch assay showed higher proliferation and migration of fibroblasts (~90-100 % wound closure) compared to electrospun PVA and control. Significant antibacterial activity was observed against Escherichia coli and Staphylococcus aureus. In vitro real-time gene expression studies with human THP-1 cell line revealed a significant downregulation of pro-inflammatory cytokines (8.64 fold decrease for TNF-α) and upregulation of anti-inflammatory cytokines (6.83 fold increase for IL-10) compared to lipopolysaccharide. In brief, the results advocate agarose-curdlan mat as a potential multifunctional, bioactive, and eco-friendly dressing for healing diabetic wounds.

Process optimization for the rapid conversion of calcite into hydroxyapatite microspheres for chromatographic applications.

Microsphere hydroxyapatite (HAp) is widely used in various biomedical and chromatographic applications. The work described in this manuscript focuses on a dissolution precipitation method for production of HAp microspheres. This method overcomes certain drawbacks of conventional preparation methods used for HAp preparation, which produce polydisperse particles and are time-consuming and expensive. In the present work, the calcium carbonate (calcite) particles were directly and rapidly converted into HAp microspheres using an inexpensive dissolution precipitation method. The effects of the reaction temperature, time, and mechanical stirring rates were studied, and the reaction parameters were optimized. As confirmed by the XRD studies, the higher reaction temperature and time promote complete HAp conversion, while calcite residues were observed for lower temperatures and times. SEM images show the influence of reaction parameters on the surface microstructure of the microspheres produced. It was observed that the HAp microspheres undergo disintegration at a higher stirring rate. The reaction parameters optimized in this work were ideal for preparing HAp microspheres. The resultant HAp particles were utilized as matrices for chromatographic separation of protein mixtures.

Fabrication and evaluation of multifunctional agarose based electrospun scaffolds for cutaneous wound repairs.

Despite several advances in chronic wound management, natural product based scaffolds with high exude absorption and mechanical strength are still a hotspot in the medical field. Thus, present study illustrates the fabrication of agarose (AG; 10% w/v)/polyvinyl alcohol 12% w/v) based multifunctional nanofibrous electrospun scaffolds. Zinc citrate (1%, 3% and 5% w/w of the polymer) was used as a potential antibacterial agent. The fabricated scaffolds exhibit a swelling of ∼550% in phosphate buffer saline and mechanical strength of 10.11 ± 0.31 MPa which is suitable for most of the wound healing applications that require high strength. In vitro study revealed an increased migration and proliferation of L929 fibroblasts with AG blends when compared to the control. The fabricated scaffolds exhibited antibacterial properties against both Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative) bacterial strains. Hence, a multifunctional (ability to protect wounds from bacterial infections along with effective swelling and mechanical support), natural product based, eco-friendly scaffold to serve as a potential wound dressing material has been successfully fabricated.

Nanocarrier-based drug delivery systems for bone cancer therapy: a review.

Bone cancer is a malignant tumor that originates in the bone and destroys the healthy bone tissues. Of the various types of bone tumors, osteosarcoma is the most commonly diagnosed primary bone malignancy. The standard treatment for primary malignant bone tumors comprises surgery, chemotherapy and radiotherapy. Owing to the lack of proven treatments, different forms of alternative therapeutic approaches have been examined in recent decades. Among the new therapeutic methodologies, nanotechnology-based anticancer therapy has paved the way for new targeted strategies for bone cancer treatment and bone regeneration. They include approaches such as the co-delivery of multiple drug cargoes, the enhancement of their biodistribution and transport properties, normalizing accumulation and the optimization of drug release profiles to overcome shortcomings of the existing therapy. This review examines the standard treatments for osteosarcoma, their lacunae, and the evolving therapeutic strategies based on nanocarrier-mediated combinational drug delivery systems, and future perspectives for osteosarcoma therapy.

Odontogenic differentiation of inflamed dental pulp stem cells (IDPSCs) on polycaprolactone (PCL) nanofiber blended with hydroxyapatite.

The objective of this study was to develop electrospun polycaprolactone (PCL) membranes blended with hydroxyapatite (HA) and evaluate its potential in differentiating inflamed dental pulp stem/progenitor cells (IDPSCs) into odontoblasts. Electrospun nanofibrous membrane consisting of PCL blended with 10 wt% and 15 wt% of HA were fabricated and the characterization was done by Scanning electron microscopy (SEM), Fourier- transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and contact angle analysis. Cytocompatibility, cell adhesion and odontogenic differentiation ability of the membranes were assessed by MTT, Live/Dead, SEM/DAPI and qPCR studies. The mineral deposition ability of the membranes with IDPSCs was estimated by SEM-EDS. The SEM analysis revealed a nanofibrous texture with an average fiber diameter of 140 nm for PCL, 220 nm for PCL10%HA and 250 nm for PCL15%HA. Among the membranes tested, PCL10%HA favored positive cell attachments, upregulated expression of DSPP and ALP gene and higher Ca/P ratio compared to PCL and PCL15%HA.

Combinational delivery of anticancer drugs for osteosarcoma treatment using electrosprayed core shell nanocarriers.

In bone cancer treatment, local delivery of chemotherapeutic agents is preferred compared to other routes of administration. Delivery of multiple drugs using biodegradable carriers improves the treatment efficiency and overcomes drug resistance and toxicity. With this approach, we have developed multilayer biodegradable core shell nanoparticles (NPs) using the electro-spraying technique to deliver methotrexate (MTX) and doxorubicin (DOX) for the treatment of osteosarcoma. These core-shell NPs with a mean particle size of 212 ± 41 nm consist of hydroxyapatite (HA) and DOX as core with the outer shell made of chitosan (CH) followed by polycaprolactone (PCL) with MTX. The encapsulation efficiency of MTX was around 85% and DOX was 38%. In vitro drug release studies were performed in phosphate buffered saline (PBS) at pH 5 and pH 7.4 for 8 days. Different release profiles were observed in both acidic and alkaline pH. The sequential release of MTX followed by DOX was observed in both pH in sustained manner. Human osteosarcoma MG 63 (OMG-63) cells lines were used to test the cytotoxicity of drug loaded NPs. Multi-drug encapsulated bioresorbable and biodegradable electro-sprayed core shell NPs will be promising as a bone substitute for the treatment of osteosarcoma.

Eggshell derived brushite bone cement with minimal inflammatory response and higher osteoconductive potential.

Brushite cements are known for excellent osteoconductive and degradation properties, however, its widespread use is limited due to rapid setting time and poor mechanical properties. The eggshell derived calcium phosphates exhibits improved physical and biological properties due to the presence of biologically relevant ions. In this study, eggshell derived brushite cement (EB) was fabricated using ß-tricalcium phosphate synthesized from eggshells. The presence of trace elements in EB prolonged its setting time. The size of brushite crystals in EB was found to be smaller than the pure brushite cement (PB) leading to increased initial compressive strength and higher in vitro degradation rate. The L6 and MG63 cell lines exhibited good biocompatibility with the cement at the end 72 h. In vivo studies of the cements were performed in rat calvarial defect model. Micro CT analysis showed faster degradation and accelerated bone formation in EB filled defect. Histological studies revealed infiltration of inflammatory cells into the implant site for both the cements till 6th week. However, inflammation was found to be significantly reduced at the 12th week in EB compared to PB leading to complete bone bridge formation. Multi-ion substituted EB seems to be a potential bone substitute material with a reasonable setting time for ease of handling, higher mechanical strength, minimal inflammatory response and higher bone regeneration.

Curcumin Releasing Eggshell Derived Carbonated Apatite Nanocarriers for Combined Anti-Cancer, Anti-Inflammatory and Bone Regenerative Therapy.

Bone cancer or osteosarcoma is an aggressive cancer affecting the long bones and is treated by a combination of surgery and chemotherapy. Local drug delivery directly to the site of bone cancer and the use of plant-based drugs has been explored towards improving the efficacy and decreasing the toxicity of the anti-cancer drugs. Curcumin, derived from turmeric is highly effective against cancer cells and shows very low toxicity against normal cells. Bone repair is facilitated by use of bone substitutes such as bioceramics, amongst which the carbonated apatite (CA) nanocarriers closely mimic the natural bone mineral. In the current work, we have developed CA nanocarriers based local delivery of curcumin as an adjunct treatment for bone cancer. CA nanocarriers with 6 wt.% carbonate were prepared by wet chemical synthesis using synthetic derived (6SWCA) and eggshell derived (6EWCA) precursors along with hydroxyapatite (WHA) as a control. The X-ray diffraction (XRD) patterns showed the CAs to be phase pure with a mean crystallite size of 17 nm. The Fouriertransform infrared spectroscopy (FTIR) analysis of both CAs indicated the carbonate substitution as B-Type. The amount of carbonate substitution was observed to be around 6 wt.% using FTIR and CHNO elemental analyzer. The 6EWCA showed a greater loading (36%) and release (66%) of curcumin than 6SWCA and WHA nanocarriers. The bovine serum albumin (BSA) protein denaturation assay showed the curcumin loaded CAs to be highly anti-inflammatory while their anti-cancer activity was confirmed by the high cytotoxic activity against MG-63 human osteosarcoma cells. Conclusively, an eggshell derived apatite drug delivery system was found to be very suitable to cure osteosarcoma, prevent post-cancer inflammation and modulate bone repair and regeneration.

Antibacterial and Bioactive Surface Modifications of Titanium Implants by PCL/TiO2 Nanocomposite Coatings.

Surface modification of biomedical implants is an established strategy to improve tissue regeneration, osseointegration and also to minimize the bacterial accumulation. In the present study, electrospun poly(ε-caprolactone)/titania (PCL/TiO2) nanocomposite coatings were developed on commercially pure titanium (cpTi) substrates for an improved biological and antibacterial properties for bone tissue engineering. TiO2 nanoparticles in various amounts (2, 5, and 7 wt %) were incorporated into a biodegradable PCL matrix to form a homogeneous solution. Further, PCL/TiO2 coatings on cpTi were obtained by electrospinning of PCL/TiO2 solution onto the substrate. The resulted coatings were structurally characterized and inspected by employing scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Given the potential biological applications of PCL/TiO2 coated cpTi substrates, the apatite-forming capacity was examined by immersing in simulated body fluid (SBF) for upto 21 days. Biocompatibility has been evaluated through adhesion/proliferation of hFOB osteoblast cell lines and cytotoxicity by MTT assay. Antimicrobial activity of PCL/TiO2 nanocomposites has been tested using UV light against gram-positive Staphylococcus aureus (S.aureus). The resulting surface displays good bioactive properties against osteoblast cell lines with increased viability of 40% at day 3 and superior antibacterial property against S.aureus with a significant reduction of bacteria to almost 76%. Surface modification by PCL/TiO2 nanocomposites makes a viable approach for improving dual properties, i.e., biological and antibacterial properties on titanium implants which might be used to prevent implant-associated infections and promoting cell attachment of orthopedic devices at the same time.

Development of Egg Shell Derived Carbonated Apatite Nanocarrier System for Drug Delivery.

Carbonated apatite has a chemical composition quite similar to biological apatite found in native bone. The incorporation of carbonate (CO2-3) ions groups into the apatitic crystal structure can tailor its crystallinity, solubility and biological activity that benefit the bone repair and regeneration. In this study, we report a simple and elegant method of synthesizing carbonated calcium deficient hydroxyapatite (ECCDHA) nanoparticles from egg shell wastes and its efficacy has been compared with synthetic calcium deficient hydroxyapatite (SCDHA) nanoparticles. Egg shell contains about 94% of calcium carbonate. Fourier transform infrared (FT-IR) spectroscopy results confirmed the carbonate substitution in the apatite as B-type and CHNS/O elemental analysis showed 6 wt.% of carbonate content in ECCDHA. Energy dispersive spectroscopy (EDS) analysis confirmed the presence of biologically relevant elements such as magnesium, strontium, fluoride, potassium etc., in ECCDHA inherited from the egg shell. In vitro cell culture studies confirmed that the ECCDHA is cellular compatible and it has enhanced cell adhesion and proliferation of L6 myoblast cells as compared to SCDHA. The potential of ECCDHA suitable for bone drug applications was tested with an antibiotic drug, doxycycline. The results showed higher drug loading and releasing for ECCDHA as compared to SCDHA during the period of study. Based on these results, the ECCDHA may be considered as a potential bone substitute and drug carrier system.

Ceramic core with polymer corona hybrid nanocarrier for the treatment of osteosarcoma with co-delivery of protein and anti-cancer drug.

For the treatment of metastatic bone cancer, local delivery of therapeutic agents is preferred compared to systemic administration. Delivery of an anti-cancer drug and a protein that helps in bone regeneration simultaneously is a challenging approach. In this study, a nanoparticulate carrier which delivers a protein and an anti-cancer drug is reported. Bovine serum albumin (BSA) as a model protein was loaded into hydroxyapatite (HA) nanoparticles (NPs) and methotrexate (MTX) conjugated to poly(vinyl alcohol) was coated onto BSA-loaded HA NPs. Coating efficiency was in the range of 10-17 wt%. In vitro drug release showed that there was a steady increase in the release of both BSA and MTX with 76% of BSA and 88% of MTX being released in 13 days. Cytotoxicity studies of the NPs performed using human osteosarcoma (OMG-63) cell line showed the NPs were highly biocompatible and exhibited anti-proliferative activity in a concentration-dependent manner.

Electrospun 3D composite scaffolds for craniofacial critical size defects.

Critical size defects in the craniofacial region can be effectively treated using three dimensional (3D) composite structures mimicking natural extra cellular matrix (ECM) and incorporated with bioactive ceramics. In this study we have shown that the dynamic liquid bath collector can be used to form electrospun polycaprolactone (PCL)-hydroxyapatite (HA) composite structure as unique 3D scaffold. The structure was found to have three distinct sections (base, stem and head) based on the mechanism of its formation and morphology. The size of the head portion was around 15 mm and was found to vary with the process parameters. Scanning electron microscopy (SEM) analysis revealed that the base had random fibres while the fibres in stem and head sections were aligned but perpendicular to each other. X-ray diffraction (XRD) analysis also showed an increase in the crystallinity index of the fibres from base to head section. Cytotoxicity and cytocompatibility studies using human osteosarcoma (HOS) cells showed good cell adhesion and proliferation indicating the suitability of the 3D structure for craniofacial graft applications.

Dentin remineralizing ability and enhanced antibacterial activity of strontium and hydroxyl ion co-releasing radiopaque hydroxyapatite cement.

Dental caries is an infection of the mineralized tooth structures that advances when acid secreted by bacterial action on dietary carbohydrates diffuses and dissolves the tooth mineral leading to demineralization. During treatment, clinicians often remove only the superficial infected tooth structures and retain a part of affected carious dentin to prevent excessive dentin loss and pulp exposure. Calcium hydroxide is used to treat the affected dentin because it is alkaline, induces pulp-dentin remineralization and decreases bacterial infection. Presence of strontium ions has also been reported to exhibit anticariogenic activity, and promote enamel and dentin remineralization. The objective of the present study was to develop novel hydroxyapatite cement from tetracalcium phosphate which gradually releases hydroxyl and strontium ions to exhibit antibacterial activity. Its potential to remineralize the dentin sections collected from extracted human molar tooth was studied in detail. The pH of all the experimental cements exhibited a gradual increase to ~10.5 in 10 days with 10% strontium substituted tetracalcium phosphate cement (10SC) showing the highest pH value which was sustained for 6 weeks. 10SC showed better antibacterial property against S. aureus and E. coli at the end of 1 week compared to other cements studied. It also exhibited the highest radiopacity equivalent to 4.8 mm of Al standard. 10SC treated dentin section showed better remineralization ability and highest elastic modulus. We can conclude that the hydroxyl and strontium ions releasing tetracalcium phosphate cement exhibits good antibacterial property, radiopacity and has the potential to encourage dentin remineralization.

Comparative evaluation of micron- and nano-sized intracanal medicaments on penetration and fracture resistance of root dentin - An in vitro study.

Intracanal medicaments play a vital role in disinfection of root canal system. The aim of this study was to evaluate intratubular penetration and fracture resistance of roots filled with micron- and nano-sized intracanal medicaments: calcium hydroxide (CH), nanocalcium hydroxide (NCH), chitosan (CS) and nanochitosan (NCS). Their antibacterial effect on E. faecalis was tested using agar diffusion method. NCH and NCS were prepared by precipitation method and ionic crosslinking respectively. NCH and NCS particles were spherical, with an average particle size of 102±11.3nm and 130±17.6nm respectively. The medicaments were filled in extracted human teeth. Depth of penetration of the medicaments into dentinal tubules at coronal (C), middle (M) and apical (A) thirds was measured. Fracture resistance of the teeth was evaluated after 1 week and 1-month intervals. NCH showed the highest depth of penetration (C ∼746.98µm, M ∼700.30µm, A ∼134.69µm). CS showed the highest fracture resistance, whereas no significant difference was found between other medicaments, at both the time intervals. NCH (8.07±0.06) and NCS (8.13±0.06) showed significantly higher zone of inhibition than CH (7.7±0.17) and CS (7.37±0.15). Under the conditions of this study, it can be concluded that NCH and NCS can be used as potential intracanal medicaments.

Electrospun Cytocompatible Polycaprolactone Blend Composite with Enhanced Wettability for Bone Tissue Engineering.

Electrospinning is recently used in tissue engineering due to their excellent ability to mimic the structure of extra cellular matrix (ECM), a prerequisite for creating an optimal microenvironment for cell growth. Electrospun nanofibrous composite scaffolds consisting of polycaprolactone (PCL)/Poly(1,4-butylene adipate-co-polycaprolactam) (PBAPCL) blend with hydroxyapatite (HA) have been fabricated to enhance the wettability and osseointegrative properties. Fourier transform-infrared spectroscopy (FT-IR) confirmed molecular interactions of the polymer blend along with the presence of HA. X-ray diffraction analysis (XRD) indicated semi-crystalline nature of the mat and also the presence of HA in the composite mat. The morphology of the fibres, were analyzed using scanning electron microscopy (SEM) and the diameter was found to be in the range of 400­600 nm. The composite fibers were of larger diameter compared to their polymer counterparts. Improved wettability of the electrospun composite mat has been observed by contact angle analysis. In vitro cell culture studies by Live/Dead assay and MTT using human osteosarcoma (HOS) cells indicated the cytocompatible nature of electrospun mat which was further confirmed by cell adhesion using SEM and Actin-phalloidin staining. Addition of PBAPCL and HA to PCL have a beneficial effect on cell growth and proliferation thereby making the composite, a prospective scaffold for bone tissue engineering applications.

Eggshell-Derived Hydroxyapatite: A New Era in Bone Regeneration.

BACKGROUND: Defects of maxillofacial skeleton lead to personal (functional and aesthetic), social and behavioral problems; which make the person to isolate from the main stream of society. So, bone regeneration is the need for proper structure, function, and aesthetics following cyst enucleation, trauma, and tumor ablative surgery; which helps for overall health of the individual. AIM AND OBJECTIVES: The preliminary study is planned to evaluate and compare the efficacy of eggshell-derived hydroxyapatite (EHA) and synthetic hydroxyapatite (SHA) following cystectomy. MATERIAL AND METHODS: Microwave-processed calcium deficient EHA and commercially available SHA are used for grafting. Total 20 patients enrolled in this study, consisting 10 in each group between 20 and 45 years of age. All the patients were evaluated for bone regeneration at first, second, third, and sixth month's interval, postsurgically, using radiovisiograph and clinical parameters. RESULTS: The bone formation characteristics vary at second month when compared to SHA. This difference may be because of the kinetics involved in the regeneration pattern. The pattern of bone healing was trabecular after third month, indicating complete bone formation. The study showed constant raise of density and remained same at the end of study period. CONCLUSIONS: Both EHA and SHA graft materials are equally efficient in early bone regeneration. Within the limitations of this study the EHA showed promising results. Which indicates the eggshell waste-bio mineral is worthwhile raw material for the production of HA and is a Go Green procedure. Eggshell-derived hydroxyapatite is economic, compared with SHA.

Eggshell derived hydroxyapatite as bone graft substitute in the healing of maxillary cystic bone defects: a preliminary report.

BACKGROUND: Since ancient times, use of graft materials to promote healing of defects of bone is wellknown. Traditionally, missing bone is replaced with material from either patient or donor. Multiple sources of bone grafts have been used to graft bone defects to stimulate bone healing. Hydroxyapatite is naturally occurring mineral component of bone, which is osteoconductive. This versatile biomaterial is derived from many sources. The aim of this study is to evaluate the efficacy of eggshell derived hydroxyapatite (EHA) in the bone regeneration of human maxillary cystic bone defects secondary to cystic removal/apicoectomy and compare the material properties of EHA in vitro. MATERIALS AND METHODS: A total of eight maxillary bone defects were grafted after cystic enucleation and/or apicoectomy in the year 2008 and completed the study at 1 year. The patients were followed-up 2 weeks after surgery for signs and symptoms of infection or any other complications that may have been related to surgical procedure. Follow-up radiographs were obtained immediately after surgery followed by 1, 2, and 3 months to assess the efficacy of EHA in bone healing. Physicochemical characterization of the EHA was carried out in comparison with synthetic hydroxyapatite (SHA), also compared the biocompatibility of EHA using in vitro cytotoxicity test. RESULTS: By the end of the 8(th) week, the defects grafted with EHA showed complete bone formation. However, bone formation in non-grafted sites was insignificant. The values of density measurements were equal or more than that of surrounding normal bone. These results indicate that the osseous regeneration of the bone defect filled with EHA is significant. EHA showed the superior material properties in comparison with SHA. CONCLUSION: EHA is a versatile novel bone graft substitute that yielded promising results. Because of its biocompatibility, lack of disease transfer risks, ease of use and unlimited availability, EHA remains a viable choice as regenerative material. EHA is very cost-effective, efficient bone graft substitute, which can be prepared in a very economical way. It is a worthwhile bone substitute because it is safe and easily available material. How to cite the article: Kattimani VS, Chakravarthi PS, Kanumuru NR, Subbarao VV, Sidharthan A, Kumar TS, Prasad LK. Eggshell derived hydroxyapatite as bone graft substitute in the healing of maxillary cystic bone defects: A preliminary report. J Int Oral Health 2014;6(3):15-9.