Alkaline Phosphate Activity Research Articles

Fabrication of chitosan-coated porous polycaprolactone/strontium-substituted bioactive glass nanocomposite scaffold for bone tissue engineering.

In the present study, porous (about 70 vol%) nanocomposite scaffolds made of polycaprolactone (PCL) and different amounts (0 to 15 wt%) of 45S bioactive glass (BG) nanoparticles (with a particle size of about 40 nm) containing 7 wt% strontium (Sr) were fabricated by solvent casting technique for bone tissue engineering. Then, a selected optimum scaffold was coated with a thin layer of chitosan containing 15 wt% Sr-substituted BG nanoparticles. Several techniques such as X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD), scanning electron microscopy (SEM), dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), tensile test, and water contact angle measurement were used to characterize the fabricated samples. In vitro experiments including degradation, bioactivity, and biocompatibility (i.e., cytotoxicity, alkaline phosphate activity, and cell adhesion) tests of the fabricated scaffold were performed. The biomedical behavior of the fabricated PCL-based composite scaffold was interpreted by considering the presence of the porosity, Sr-substituted BG nanoparticles, and the chitosan coating. In conclusion, the fabricated chitosan-coated porous PCL/BG nanocomposite containing 15 wt% BG nanoparticles could be utilized as a good candidate for bone tissue engineering.

Functionally graded titanium implants: Characteristic enhancement induced by combined severe plastic deformation.

Commercially pure titanium was processed by equal channel angular pressing (ECAP) and surface mechanical attrition treatment (SMAT) for the purpose of developing functionally graded titanium used for implants and a gradient structure including nanostructured, deformed and undeformed zones were produced on the samples. In particular, it was aimed to design the gradient-structure in the titanium with enhanced properties by applying 4 ECAP passes to form bulk structure of ultrafine-grains and subsequently subjecting SMAT to the surface of ECAPed samples to produce nanostructured surface region. Microstructural examination was made by electron back scatter diffraction (EBSD). Also, microhardness, nanoindentation, topography, roughness and wettability were evaluated. To examine the biological response, human osteosarcoma cells were cultured in contact with the samples in various time periods and morphology change, cell viability and alkaline phosphate activity were conducted also cell morphology was monitored. EBSD showed development of ultrafine-grained structure after 4 passes of ECAP with an average grain size of 500 nm. Applying SMAT resulted in additional refinement in the ECAP samples, particularly in the subsurface regions to a depth of 112 μm. Furthermore, the SMATed samples showed an enhancement in roughness, wettability and hardness magnitudes. Viability enhanced up to 7% in SMATed + ECAPed sample, although the acceptable cell adhesion, improved cell differentiation and mineralization were seen. The combined use of ECAP and SMAT has shown a good potential for optimizing the design of modern functionally graded medical devices and implants.

The Synergic Effect of Terpenoid and Steroidal Saponins Can Improve Bone Healing, by Promoting the Osteogenic Commitment of Adipose Mesenchymal Stem Cells: An In Vitro Study

Bone regeneration involves several biological processes that consistently impact the quality of tissue healing. An important step consists of the local recruitment and differentiation of mesenchymal stem cells that migrate in the site to regenerate from bone marrow. Mesenchymal stem cells (MSCs) may be pushed towards osteogenic commitment by specific substances, often naturally present in plants. Yunnan Baiyao (YB) is a Chinese herbal medicine, mainly working through the synergic effect of terpenoid and steroidal saponins. YB is well known for its numerous biomedical effects, including the ability to favor improved bone tissue healing. In our in vitro study, we used adipose mesenchymal stem cells (ADSCs) as a study-model: We selected samples to harvest and isolate ADSCs and investigate their viability; moreover, we performed bone-related gene expression to evaluate the differentiation of MSCs. To confirm this behavior, we analyzed alkaline phosphate activity and calcium deposition, with ADSCs cultured in basal and osteogenic media, with YB at different concentrations in the medium, and at different time-points: 7, 14 and 21 days. Our results indicate that the synergic effect of terpenoid and steroidal saponins slightly favor the late ADSCs differentiation towards the osteoblasts phenotype. In osteogenic committed cells, the treatment with the lower dose of YB promoted the up-regulation of the alkaline phosphatase gene (ALPL) at day seven and 14 (p < 0.01); at day 21, the alkaline phosphatase (ALP) activity showed a slight increase, although in basal condition it maintains low rates. We assume that such molecular synergy can promote the osteogenic commitment of adipose mesenchymal stem cells, thus improving the timing and the quality of bone healing.

Cyanobacteria in eutrophic waters benefit from rising atmospheric CO2 concentrations.

Rising atmospheric carbon dioxide (CO2) may stimulate the proliferation of cyanobacteria. To investigate the possible physiological responses of cyanobacteria to elevated CO2 at different nutrient levels, Microcystis aeruginosa were exposed to different concentrations of CO2 (400, 1100, and 2200 ppm) under two nutrient regimes (i.e., in nutrient-rich and nutrient-poor media). The results indicated that M. aeruginosa differed in its responses to elevated atmospheric CO2 at different nutrient levels. The light utilization efficiency and photoprotection of photosystem II were improved by elevated CO2, particularly when cells were supplied with abundant nutrients. In nutrient-poor media, both total organic carbon and the polysaccharide/protein ratio of the extracellular polymeric substance increased with elevated CO2, accompanied by high cellular carbon/nitrogen ratios. Besides, cells growing with fewer nutrients were more prone to suffer intracellular acidification with elevated CO2 than those growing with abundant nutrients. Nonetheless, alkaline phosphate activity of cyanobacteria was improved by high CO2, provided that reduced pH was in the optimum range for alkaline phosphate activity. Nitrate reductase activity was inhibited by elevated CO2 regardless of nutrient levels, leading to a reduced nitrate uptake. These changes indicate that the biogeochemical cycling of nutrients would be affected by higher atmospheric CO2 conditions. Overall, cyanobacteria in eutrophic waters may benefit more than in oligotrophic waters from rising atmospheric CO2 concentrations, and evaluations of the influence of rising atmospheric CO2 on algae should account for the nutrient level of the ecosystem.

Dual effective core-shell electrospun scaffolds: Promoting osteoblast maturation and reducing bacteria activity

Herein, we electrospun ultrathin core-shell fibers based on polycaprolactone (PCL), polyethylene glycol (PEG), gelatin and osteogenic growth peptide (OGP), and evaluated their potential to upregulate human osteoblast cells (hFOB) and to reduce Gram-positive and Gram-negative bacteria. We also evaluated the fiber morphology, chemical structure and peptide delivery efficacy. The employment of core-shell fibers compared to fibers without a core-shell showed improved mechanical strength, comparable to the strength of pure PCL, as well as improved hydrophilicity and wettability. The careful selection of polymer combination and core-shell strategy promoted a controlled and sustained release of OGP. Moreover, increased calcium deposition (CD) (1.3-fold) and alkaline phosphate (ALP) activity was observed when hFOBs were cultivated onto core-shell fibers loaded with OGP after 21 days of culture. Our developed scaffolds were also able to reduce the amount of Pseudomonas aeruginosa (ATCC 25668) bacteria by a factor of two compared to raw PCL without the use of any antibiotics. All of these results demonstrate a promising potential of the developed core-shell electrospun scaffolds based on PCL:PEG:Gelatin:OGP for numerous bone tissue applications.

Toxicological Safety Assessment of Molluscicides Against Non-target Aquatic Biota; Colisa fasciatus

Plants Terminalia arjuna and Tamarindus indica are known to have a significant molluscicidal potential to control the population of vector snails. Since the molluscicides are considered an emerging pollutant and are frequently detected in surface water bodies and found to be a great threat for aquatic biota. Hence, the main objective of this research is to critically evaluate the ectotoxicological and chronic effect of plant-derived molluscicides on other aquatic biotas. With these references this study deals with the safety measurement of molluscicides arjnolic acid, saponin and procynadine from T. arjuna and T. Indica against the fish Colisa fasciatus which share the same habitat with snails. The result of toxicity experiment reveals that fishes showed no mortality against 24h LC90 (against L. acuminata and I. exustus) up to 96h exposure duration. The enzyme bioassays of these molluscicides on the nervous tissue of fish showed no significant effect on key enzymes Acetylcholinesterase, acid and alkaline phosphates activity in comparison to control group of fishes. These results indicated that the application of arjunolic acid, saponin, and procynadine derive from plant T. arjuna and T. indica at its maximum concentration (24h LC90 of L. acuminata and I. exustus) and exposure duration (96h) did not cause any mortality or treatment-related enzymes inactivity in fishes. The study conclusively proved the ecotoxicological and chronic safety of plant-derived molluscicides arjunolic acid, saponin, and procynadine on non-target animals in the aquatic environment.

Enhanced cell functions on graphene oxide incorporated 3D printed polycaprolactone scaffolds

For tissue engineering applications, a porous scaffold with an interconnected network is essential to facilitate the cell attachment and proliferation in a three dimensional (3D) structure. This study aimed to fabricate the scaffolds by an extrusion-based 3D printer using a blend of polycaprolactone (PCL), and graphene oxide (GO) as a favorable platform for bone tissue engineering. The mechanical properties, morphology, biocompatibility, and biological activities such as cell proliferation and differentiation were studied concerning the two different pore sizes; 400 μm, and 800 μm, and also with two different GO content; 0.1% (w/w) and 0.5% (w/w). The compressive strength of the scaffolds was not significantly changed due to the small amount of GO, but, as expected scaffolds with 400 μm pores showed a higher compressive modulus in comparison to the scaffolds with 800 μm pores. The data indicated that the cell attachment and proliferation were increased by adding a small amount of GO. According to the results, pore size did not play a significant role in cell proliferation and differentiation. Alkaline Phosphate (ALP) activity assay further confirmed that the GO increase the ALP activity and further Elemental analysis of Calcium and Phosphorous showed that the GO increased the mineralization compared to PCL only scaffolds. Western blot analysis showed the porous structure facilitate the secretion of bone morphogenic protein-2 (BMP-2) and osteopontin at both day 7 and 14 which galvanizes the osteogenic capability of PCL and PCL + GO scaffolds.

Comparison of osteogenic differentiation potential of induced pluripotent stem cells on 2D and 3D polyvinylidene fluoride scaffolds.

In recent decades, tissue engineering has been the most contributor for introducing 2D and 3D biocompatible osteoinductive scaffolds as bone implants. Polyvinylidene fluoride (PVDF), due to the unique mechanical strength and piezoelectric properties, can be a good choice for making a bone bioimplant. In the present study, PVDF nanofibers and film were fabricated as 3D and 2D scaffolds, and then, osteogenic differentiation potential of the human induced pluripotent stem cells (iPSCs) was investigated when grown on the scaffolds by evaluating the common osteogenic markers in comparison with tissue culture plate. Biocompatibility of the fabricated scaffolds was confirmed qualitatively and quantitatively by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and scanning electron microscopy assays. Human iPSCs cultured on PVDF nanofibers showed a significantly higher alkaline phosphate activity and calcium content compared with the iPSCs cultured on PVDF film. Osteogenic-related genes and proteins were also expressed in the iPSCs seeded on PVDF nanofibers significantly higher than iPSCs seeded on PVDF film, when investigated by real-time reverse transcription polymerase chain reaction and western blot analysis, respectively. According to the results, the PVDF nanofibrous scaffold showed a greater osteoinductive property compared with the PVDF film and due to the material similarity of the scaffolds, it could be concluded that the 3D structure could lead to better bone differentiation. Taken together, the obtained results demonstrated that human iPSC-seeded PVDF nanofibrous scaffold could be considered as a promising candidate for use in bone tissue engineering applications.

SiO2 loading into polydopamine-functionalized TiO2 nanotubes for biomedical applications

A recent advancement in bone therapies is the development of advanced implants from arrays of titania nanotubes, which are fabricated with self-ordering electrochemical anodization of a Ti substrate. We have fabricated a novel interface by coating an anodized Ti substrate with polydopamine. This interface is suitable for the efficient incorporation of SiO2 nanoparticles into the porous layer of TiO2 nanotubes formed on the underlying Ti substrate. The results of different characterization tests indicate that the nanoparticles are evenly incorporated into the titania nanotube arrays. Potentiodynamic polarization curves and electrochemical impedance spectra measurements in a solution of simulated body-fluid indicate that the composite layer of SiO2 nanoparticles and the titania-nanotube arrays were more corrosive-resistant than the layer of titania-nanotubes arrays. Its hydrophilic properties and surface roughness are enhanced with the incorporation of SiO2 nanoparticles. As shown by in-vitro cellular assays, cell attachment and alkaline phosphate activity on such composite layer improved. The polydopamine/anodization treatment of Ti substrates allows the quick integration of SiO2 with titania nanotubes to increase corrosive resistance and biocompatibility of the Ti substrate. This could be a new approach to investigating the collective effects of local chemistry of compounds, e.g., SiO2, and of local topography of titania nanotubes.

Experimental study on effect of Naga Bhasma (lead calx) on hematological and biochemical parameters

Background: In recent decade, a lot of concerns has been raised regarding heavy metal content in Ayurveda medicines, especially Bhasma (metallic calx). Naga Bhasma (lead calx) is of the metallic preparation primarily indicated in treating chronic ailments such as Kamala (jaundice), Vatarakta (gout), Grdhrasi (sciatica), Kustha (skin diseases), and Prameha (diabetes). As being a compound of lead, its chronic toxicity study was planned to find out the effect of Naga Bhasma on serum biochemical and hematological parameters. Material and method: Naga Bhasma was prepared by Parada (mercury) and Gandhaka media (NBP) and Vasa Swarasa (juice of Adhatoda vasica Linn.) as herbal media (NBH) in seven Puta (incineration cycles). Chronic oral toxicity study was carried out as per Organization for Economic cooperation and Development (OECD) 408 (90-day oral repeated dose toxicity study). Wistar strain albino rats (Rattus norvegicus) were used for the experimentation. The study was conducted at three dosage levels, therapeutically effective dose (TED) of NBP and NBH was 45 mg/kg body weight of rat, TED × 5 and TED × 10. On 91st day, blood was collected by supraorbital puncture with the help of microcapillary tubes under mild ether anesthesia for estimation of serum biochemical and hematological parameters. Observation and results: Among hematological parameters, significant decrease was observed in MCV (Mean corpuscular volume), MCH (Mean corpuscular haemoglobin), and mean corpuscular hemoglobin concentration by NBH-treated group, whereas in biochemical parameters, both test drugs significantly increased BSL (Blood Sugar Level), SGOT (Serum Glutamic-Oxaloacetic Transaminase), and serum ALP (Alkaline Phosphatise) activity. Hematological and biochemical analysis indicates that both test drugs (prepared in seven incineration cycles) were not found completely safe, especially at higher dosage where the drug affected renal and hepatic functions. Conclusion: For the safety purpose, it is suggested that Naga Bhasma prepared in less number of incineration cycles may not be used for therapeutic purpose.

Tricalcium silicate/graphene oxide bone cement with photothermal properties for tumor ablation.

Bone cements have been used in the clinical setting to fill bone defects resulting from bone tumors. However, traditional bone cements do not have the function to kill tumor cells. This study develops a new type of tricalcium silicate (CS) based functional bone cement with excellent photothermal performance for the minimally invasive therapy of bone defects as well as bone tumors. Graphene oxide (GO) was introduced into the CS cement by co-precipitation of CS particles with GO nanosheets to form a CS/GO composite material based on charge interactions between the CS and GO. The incorporation of GO enhanced the self-setting properties of CS and endowed the cement with excellent photothermal performance with the irradiation of near-infrared light. Besides this, the temperature of the composite cement could be regulated by adjusting the laser power and the GO content, where the rising temperature significantly inhibited the growth of subcutaneous tumor tissue in vivo. In addition, the hydration process and development of the early compressive strength of the composite cement could be modulated based on its photothermal performance. Moreover, the CS/GO composite cement retained the bioactivity of CS to promote cell proliferation and the alkaline phosphate activity of MC3T3-E1. Therefore, the CS/GO composite cement holds great promise as a new type of functional bone cement with photothermal performance for bone tumor therapy and bone defect repair.

The responses of soil microbial community and enzyme activities of Phoebe zhennan cultivated under different soil moisture conditions to phosphorus addition

The importance of conservation and ecological restoration of the rare and economically important tree Phoebe zhennan is increasingly recognized. To this purpose, phosphorus (P) addition has been proposed to improve soil biological attributes and face the anticipated drought under climate change, though few studies have investigated its effect on the interaction between the soil microorganisms and plant host, as well as on ecosystem productivity. We investigated the effect of P addition on soil chemical properties, microbial communities, and enzyme activities in a soil planted with P. zhennan under two levels of water treatments (optimum water and drought treatments). P additions had no significant effect on microbial communities, dissolved organic nitrogen (DON), pH and soil moisture (SM), though the available P (aP) increased. Compared with no P treatment, alkaline phosphate and β-fructofuranosidase activities increased with P additions in the drought treatment. Drought decreased the total phospholipid-derived fatty acids (PLFAs), arbuscular mycorrhiza fungi (AMF), and fungi PLFAs compared to the well-watered. These findings indicated that P additions does not ameliorate the impact of drought on soil microbial communities and enzyme activities, except alkaline phosphate and β-fructofuranosidase, and P may not be responsible for regulating biochemical processes essential for maintaining the fertility of soil planted with P. zhennan under drought conditions. It is hypothesized that the lack of effects of P addition on the majority of the microbial properties could be due to the soil mechanism employed by P. zhennan to tolerate harsh conditions.

Markers of Liver function in rabbits (Oryctolagus cuniculus): Bark extracts of Nauclea latifolia and Alchornea cordifolia plants

Introduction: The use of Nauclea latifolia and Alchornea cordifolia plants as herbal curative medicine is fast growing and the need to investigate the likely toxicity to avert severe medical issues. Aims: To assess the toxicity level, the liver function enzyme activities in rabbits (Oryctolagus cuniculus) administered with both ethanolic bark extracts of Nauclea latifolia and Alchornea cordifolia were studied. Materials and Methods: Eighty-four (84) rabbits were arbitrarily grouped into seven (n = 12), where group 1 is the control, and the remaining groups were orally administered with 500, 750 and 1000 mg/kg body weight of ethanolic bark extract of Nauclea latifolia and Alchornea cordifolia (LD50>1000mg/kg). In each group, after 24 hours of administration on the 1st, 3rd and 7th days, three animals were sacrificed. Markers of liver functions: serum alkaline phosphate (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzymes activities were monitored using spectrophotometry instrument. Results: The results show a significant (p < 0.05) reduction of ALP, AST and ALT activities at all doses for the days of administration of both ethanolic bark extracts of Nauclea latifolia and Alchornea cordifolia when compared to the control. Conclusion: The study suggests that ethanolic bark extracts of Nauclea latifolia and Alchornea cordifolia may possess relative hepatoprotective activity.

HDAC6 regulates dental mesenchymal stem cells and osteoclast differentiation

BackgroundDental and periodontal tissue development is a complicated process involving a finely regulated network of communication among various cell types. Understanding the mechanisms involved in regulating dental mesenchymal stem cells (MSCs) and osteoclast cell differentiation is critical. However, it is still unclear whether histone deacetylase HDAC6 is involved in dental MSCs fate determination and osteoclast differentiation.MethodsWe used shRNA and siRNA knockdown to explore the role of HDAC6 in dental MSCs odontogenic differentiation and osteoclasts maturation.ResultsBased on HDAC6 knockdown dental MSCs, our data suggest that HDAC6 knockdown significantly increases alkaline phosphate activity and mineralized nodules formation. Additionally, mRNA expression of odontogenic marker genes (OSX, OCN, and OPN) was induced by HDAC6 knockdown. By using HDAC6 siRNA, we knocked down HDAC6 in osteoclast precursor RAW 264.7 cells. Our data suggests that HDAC6 knockdown significantly inhibited osteoclasts differentiation. Additionally, mRNA expression of osteoclast marker genes Trap, Mmp9, and Ctsk was decreased by HDAC6 knockdown.ConclusionsOur study demonstrated that HDAC6 plays an important role in regulating dental MSCs and osteoclasts differentiation.

Topographically Defined, Biodegradable Nanopatterned Patches to Regulate Cell Fate and Acceleration of Bone Regeneration.

If only allowed to proceed naturally, the bone-healing process can take several weeks, months, or even years depending on the injury size. In terms of bone-healing speed, many studies have been conducted investigating the deliverance of various growth factors of implantable biomaterials to shorten the time for bone regeneration. However, there may be side effects such as nerve pain, infection, or ectopic bone formation. As an alternative method, we focused on biophysical guidance, which provided similar topographical cues to the cellular environment to recruit host cells for bone defect healing. In this study, we hypothesized that aligned nanotopographical features have enhanced osteoblast recruitment, migration, and differentiation without external stimuli. We designed and fabricated a biodegradable poly(lactic- co-glycolic acid) nanopatterned patch using simple solvent casting and capillary force lithography. We confirmed that a biodegradable nanopatterned patch (BNP) accelerated the migration of osteoblasts according to the orientation of the patterned direction. These highly aligned osteoblasts may contribute to in vitro osteogenic differentiation, such as alkaline phosphate activity, mineralization, and calcium deposition, compared to the biodegradable flat patch (BFP). To demonstrate bone defect healing by BNP guidance in vivo, we implanted either whole or bridge BNP on the critical size defect of mouse calvarial ( ø 4 mm) or tibia bone (3 × 7 mm2). Only the BNP-treated group showed faster new bone formation and compact bone regeneration at the calvarial or tibia bone defect area compared to BFP at 4 or 8 weeks. Bridge BNP guided, in particular, the regeneration of new bone formation along the parallel direction of nanopatterned substrates. Here, we show that a BNP with biophysical guidance should be suitable for use in bone tissue regeneration through accelerated migration of the intact host cell.

Adrenocortical Challenge Response and Genomic Analyses in Scottish Terriers With Increased Alkaline Phosphate Activity.

Scottish terriers (ST) frequently have increased serum alkaline phosphatase (ALP) of the steroid isoform. Many of these also have high serum concentrations of adrenal sex steroids. The study's objective was to determine the cause of increased sex steroids in ST with increased ALP. Adrenal gland suppression and stimulation were compared by low dose dexamethasone (LDDS), human chorionic gonadotropin (HCG) and adrenocorticotropic hormone (ACTH) response tests. Resting plasma pituitary hormones were measured. Steroidogenesis-related mRNA expression was evaluated in six ST with increased ALP, eight dogs of other breeds with pituitary-dependent hyperadrenocorticism (HAC), and seven normal dogs. The genome-wide association of single nucleotide polymorphisms (SNP) with ALP activity was evaluated in 168 ST. ALP (reference interval 8–70 U/L) was high in all ST (1,054 U/L) and HAC (985 U/L) dogs. All HAC dogs and 2/8 ST had increased cortisol post-ACTH administration. All ST and 2/7 Normal dogs had increased sex steroids post-ACTH. ST and Normal dogs had similar post-challenge adrenal steroid profiles following LDDS and HCG. Surprisingly, mRNA of hydroxysteroid 17-beta dehydrogenase 2 (HSD17B2) was lower in ST and Normal dogs than HAC. HSD17B2 facilities metabolism of sex steroids. A SNP region was identified on chromosome 5 in proximity to HSD17B2 that correlated with increased serum ALP. ST in this study with increased ALP had a normal pituitary-adrenal axis in relationship to glucocorticoids and luteinizing hormone. We speculate the identified SNP and HSD17B2 gene may have a role in the pathogenesis of elevated sex steroids and ALP in ST.

Enhanced Skull Bone Regeneration by Sustained Release of BMP-2 in Interpenetrating Composite Hydrogels.

Direct administration of bone morphogenetic protein-2 (BMP-2) for bone regeneration could cause various clinical side effects such as osteoclast activation, inflammation, adipogenesis, and bone cyst formation. In this study, thiolated gelatin/poly(ethylene glycol) diacrylate (PEGDA) interpenetrating (IPN) composite hydrogels were developed for guided skull bone regeneration. To promote bone regeneration, either polycation-based coacervates (Coa) or gelatin microparticles (GMPs) were incorporated within IPN gels as BMP-2 carriers. Both BMP-2 loaded Coa and BMP-2 loaded GMPs showed significantly enhanced in vitro alkaline phosphate (ALP) activity of human mesenchymal stem cells (hMSCs) than non-BMP-2 treated control. Moreover, BMP-2 loaded GMPs group exhibited statistically increased ALP activity compared to both bolus BMP-2 administration and BMP-2 loaded Coa group, indicating that our carriers could protect and maintain biological activity of cargo BMP-2. Sustained release kinetics of BMP-2 from IPN composite hydrogels could be controlled by different formulations. For in vivo bone regeneration, various IPN gel formulations (i.e., (1) control, (2) only hydrogel, (3) hydrogel with bolus BMP-2, (4) hydrogel with BMP-2-loaded Coa, and (5) hydrogel with BMP-2-loaded GMPs) were bilaterally implanted into 5 mm-sized rat calvarial defects. After 4 weeks, micro-CT and histological analysis were performed to evaluate new bone formation. Significantly higher scores for bony bridging and union were observed in BMP-2-loaded Coa and BMP-2-loaded GMP groups as compared to other formulations. In addition, rats treated with BMP-2-loaded GMPs showed a significantly higher ratio of bone volume/total volume and lower trabecular separation scores than others. Finally, rats treated with either Coa or GMP groups exhibited a significant increase in bone formation area, as assessed via histomorphometric analysis. Taken together, it could be concluded that Coa and GMPs were effective carriers to maintain the bioactivity of cargo BMP-2 during its sustained release. Consequently, our IPN composite hydrogel system that combines such BMP-2 carriers could effectively promote skull bone regeneration.

Combinatorial Screening of Nanoclay-Reinforced Hydrogels: A Glimpse of the "Holy Grail" in Orthopedic Stem Cell Therapy?

Despite the promise of hydrogel-based stem cell therapies in orthopedics, a significant need still exists for the development of injectable microenvironments capable of utilizing the regenerative potential of donor cells. Indeed, the quest for biomaterials that can direct stem cells into bone without the need of external factors has been the "Holy Grail" in orthopedic stem cell therapy for decades. To address this challenge, we have utilized a combinatorial approach to screen over 63 nanoengineered hydrogels made from alginate, hyaluronic acid, and two-dimensional nanoclays. Out of these combinations, we have identified a biomaterial that can promote osteogenesis in the absence of well-established differentiation factorssuch as bone morphogenetic protein 2 (BMP2) or dexamethasone. Notably, in our "hit" formulationswe observed a 36-fold increase in alkaline phosphate (ALP)activity and a 11-fold increase in the formation of mineralized matrix, compared to the control hydrogel. This induced osteogenesis was further supported by X-ray diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive X-ray spectroscopy. Additionally, theMontmorillonite-reinforced hydrogels exhibited high osteointegration as evident from the relatively stronger adhesion to the bone explants as compared to the control. Overall, our results demonstrate the capability of combinatorial and nanoengineered biomaterials to induce bone regeneration through osteoinduction of stem cells in a natural and differentiation-factor-free environment.

Enhancement of osteogenic differentiation of adipose-derived stem cells by PRP modified nanofibrous scaffold.

Recent developments in bone tissue engineering have paved the way for more efficient and cost-effective strategies. Additionally, utilization of autologous sources has been considered very desirable and is increasingly growing. Recently, activated platelet rich plasma (PRP) has been widely used in the field of bone tissue engineering, since it harbours a huge number of growth factors that can enhance osteogenesis and bone regeneration. In the present study, the osteogenic effects of PRP coated nanofibrous PES/PVA scaffolds on adipose-derived mesenchymal stem cells have been investigated. Common osteogenic markers were assayed by real time PCR. Alkaline phosphate activity, calcium deposition and Alizarin red staining assays were performed as well. The results revealed that the highest osteogenic differentiation occurred when cells were cultured on PRP coated PES/PVA scaffolds. Interestingly, direct application of PRP to culture media had no additive effects on osteogenesis of cells cultured on PRP coated PES/PVA scaffolds or those receiving typical osteogenic factors. The highest osteogenic effects were achieved by the simplest and most cost-effective method, i.e. merely by using PRP coated scaffolds. PRP coated PES/PVA scaffoldscan maximally induce osteogenesis with no need for extrinsic factors. The major contribution of this paper to the current researches on bone regeneration is to suggest an easy, cost-effective approach to enhance osteogenesis via PRP coated scaffolds, with no additional external growth factors.

The effect of human-like collagen calcium complex on osteoporosis mice

The objective of this study was to assess the effect of a modified human-like collagen calcium complex on osteoporosis mice. BHK (Baby Hamster Kidney) cells were used to compare the cytotoxicity of different calcium reagents with the MTT test. Six-week-old male mice (n = 80) were randomly divided into eight groups: a blank group (blank), control group (control), human-like collagen calcium group (HLC-Ca), thiolated human-like collagen calcium group (SH-HLC-Ca), phosphorylated human-like collagen calcium group (Pi-HLC-Ca), gluconate group (Glc-Ca), calcium carbonate group (CaCO3) and D-cal group (B). A systematic analysis of the results available in vivo after 3 months of treatment was presented. The effects of several Ca supplements on osteoporosis mice were investigated by detecting serum calcium, alkaline phosphate activity (ALP), bone hydroxyproline (BHP) and bone mineral density (BMD). The results proved that the BMD and BHP of osteoporosis mice were significantly increased in the Pi-HLC-Ca group, while serum calcium and ALP were decreased. Therefore, Pi-HLC-Ca is likely a good calcium supplement for clinical applications. In this review, the advantage of Pi-HLC-Ca in preventing and delaying osteoporosis is highlighted. In addition to the current progress, further investigations are necessary to reveal the relative influences of collagen and calcium proportions on the long-term clinical effects of osteoporosis.