Biomechanics of circulating cellular and subcellular bioparticles: beyond separation.

Biomechanical attributes have emerged as novel markers, providing a reliable means to characterize cellular and subcellular fractions. Numerous studies have identified correlations between these factors and patients' medical status. However, the absence of a thorough overview impedes their applicability in contemporary state-of-the-art therapeutic strategies. In this context, we provide a comprehensive analysis of the dimensions, configuration, rigidity, density, and electrical characteristics of normal and abnormal circulating cells. Subsequently, the discussion broadens to encompass subcellular bioparticles, such as extracellular vesicles (EVs) enriched either from blood cells or other tissues. Notably, cell sizes vary significantly, from 2 µm for platelets to 25 µm for circulating tumor cells (CTCs), enabling the development of size-based separation techniques, such as microfiltration, for specific diagnostic and therapeutic applications. Although cellular density is relatively constant among different circulating bioparticles, it allows for reliable density gradient centrifugation to isolate cells without altering their native state. Additionally, variations in EV surface charges (-6.3 to -45 mV) offer opportunities for electrophoretic and electrostatic separation methods. The distinctive mechanical properties of abnormal cells, compared to their normal counterparts, present an exceptional opportunity for diverse medical and biotechnological approaches. This review also aims to provide a holistic view of the current understanding of popular techniques in this domain that transcend conventional boundaries, focusing on early harvesting of malignant cells from body fluids, designing effective therapeutic options, cell targeting, and resonating with tissue and genetic engineering principles.

Fabrication and characterization of 3D-printed composite scaffolds of coral-derived hydroxyapatite nanoparticles/polycaprolactone/gelatin carrying doxorubicin for bone tissue engineering.

In this study, nanocomposite scaffolds of hydroxyapatite (HA)/polycaprolactone (PCL)/gelatin (Gel) with varying amounts of HA (42-52 wt. %), PCL (42-52 wt. %), and Gel (6 wt. %) were 3D printed. Subsequently, a scaffold with optimal mechanical properties was utilized as a carrier for doxorubicin (DOX) in the treatment of bone cancer. For this purpose, HA nanoparticles were first synthesized by the hydrothermal conversion of Acropora coral and characterized by using different techniques. Also, a compression test was performed to investigate the mechanical properties of the fabricated scaffolds. The mineralization of the optimal scaffold was determined by immersing it in simulated body fluid (SBF) solution for 28 days, and the biocompatibility was investigated by seeding MG-63 osteoblast-like cells on it after 1-7 days. The obtained results showed that the average size of the synthesized HA particles was about 80 nm. The compressive modulus and strength of the scaffold with 47 wt. % HA was reported to be 0.29 GPa and 9.9 MPa, respectively, which was in the range of trabecular bones. In addition, the scaffold surface was entirely coated with an apatite layer after 28 days of soaking in SBF. Also, the efficiency and loading percentage of DOX were obtained as 30.8 and 1.6%, respectively. The drug release behavior was stable for 14 days. Cytotoxicity and adhesion evaluations showed that the fabricated scaffold had no negative effects on the viability of MG-63 cells and led to their proliferation during the investigated period. From these results, it can be concluded that the HA/PCL/Gel scaffold prepared in this study, in addition to its drug release capability, has good bioactivity, mechanical properties, and biocompatibility, and can be considered a suitable option for bone tumor treatment.

In vivo evaluation of bone regeneration using ZIF8-modified polypropylene membrane in rat calvarium defects.

AIM: The profound potential of zeolitic imidazolate framework 8 (ZIF8) thin film for inducing osteogenesis has been previously established under in vitro conditions. As the next step towards the clinical application of ZIF8-modified substrates in periodontology, this in vivo study aimed to evaluate the ability of the ZIF8 crystalline layer to induce bone regeneration in an animal model defect. MATERIALS AND METHODS: Following the mechanical characterization of the membranes and analysing the in vitro degradation of the ZIF8 layer, in vivo bone regeneration was evaluated in a critical-sized (5-mm) rat calvarial bone defect model. For each animal, one defect was randomly covered with either a polypropylene (PP) or a ZIF8-modified membrane (n = 7 per group), while the other defect was left untreated as a control. Eight weeks post surgery, bone formation was assessed by microcomputed tomography scanning, haematoxylin and eosin staining and immunohistochemical analysis. RESULTS: The ZIF8-modified membrane outperformed the PP membrane in terms of mechanical properties and revealed a trace Zn+2 release. Results of in vivo evaluation verified the superior barrier function of the ZIF8-coated membrane compared with pristine PP membrane. Compared with the limited marginal bone formation in the control and PP groups, the defect area was almost filled with mature bone in the ZIF8-coated membrane group. CONCLUSIONS: Our results support the effectiveness of the ZIF8-coated membrane as a promising material for improving clinical outcomes of guided bone regeneration procedures, without using biological components.

Potential neuroprotective effect of stem cells from apical papilla derived extracellular vesicles enriched by lab-on-chip approach during retinal degeneration.

Retinal degeneration (RD) is recognized as a frequent cause of visual impairments, including inherited (Retinitis pigmentosa) and degenerative (age-related macular) eye diseases. Dental stem cells (DSCs) have recently demonstrated a promising neuroprotection potential for ocular diseases through a paracrine manner carried out by extracellular vesicles (EVs). However, effective isolation of EVs is still challenging, and isolation methods determine the composition of enriched EVs and the subsequent biological and functional effects. In the present study, we assessed two enrichment methods (micro-electromechanical systems and ultrafiltration) to isolate the EVs from stem cells from apical papilla (SCAP). The size distribution of the corresponding isolates exhibited the capability of each method to enrich different subsets of EVs, which significantly impacts their biological and functional effects. We confirmed the neuroprotection and anti-inflammatory capacity of the SCAP-EVs in vitro. Further experiments revealed the possible therapeutic effects of subretinal injection of SCAP-EVs in the Royal College of Surgeons (RCS) rat model. We found that EVs enriched by the micro-electromechanical-based device (MEMS-EVs) preserved visual function, reduced retinal cell apoptosis, and prevented thinning of the outer nuclear layer (ONL). Interestingly, the effect of MEMS-EVs was extended to the retinal ganglion cell/retinal nerve fiber layer (GCL/RNFL). This study supports the use of the microfluidics approach to enrich valuable subsets of EVs, together with the choice of SCAP as a source to derive EVs for cell-free therapy of RD.

AC electrokinetic isolation and detection of extracellular vesicles from dental pulp stem cells: Theoretical simulation incorporating fluid mechanics.

Extracellular vesicles (EVs) are cell-derived nanoscale vesicles involved in intracellular communication and the transportation of biomarkers. EVs released by mesenchymal stem cells have been recently reported to play a role in cell-free therapy of many diseases. However, the demand for better research tools to replace the tedious conventional methods used to study EVs is getting stronger. EVs' manipulation using alternating current (AC) electrokinetic forces in a microfluidic device has appeared to be a reliable and sensitive diagnosis and trapping technique. Given that different AC electrokinetic forces may contribute to the overall motion of particles and fluids in a microfluidic device, EVs' electrokinetic trapping must be examined considering all dominant forces involved depending on the experimental conditions. In this paper, AC electrokinetic trapping of EVs using an interdigitated electrode arrays is investigated. A 2D numerical simulation incorporating the two significant AC electrokinetic phenomena (Dielectrophoresis and AC electroosmosis) has been performed. Theoretical predictions are then compared with experimental results and allow for a plausible explanation of observations inconsistent with DEP theory. It is demonstrated that the inconsistencies can be attributed to a significant extent to the contribution of the AC electroosmotic effect.

Recent developments in enzyme immobilization technology for high-throughput processing in food industries.

The demand for food and beverage markets has increased as a result of population increase and in view of health awareness. The quality of products from food processing industry has to be improved economically by incorporating greener methodologies that enhances the safety and shelf life via the enzymes application while maintaining the essential nutritional qualities. The utilization of enzymes is rendered more favorable in industrial practices via the modification of their characteristics as attested by studies on enzyme immobilization pertaining to different stages of food and beverage processing; these studies have enhanced the catalytic activity, stability of enzymes and lowered the overall cost. However, the harsh conditions of industrial processes continue to increase the propensity of enzyme destabilization thus shortening their industrial lifespan namely enzyme leaching, recoverability, uncontrollable orientation and the lack of a general procedure. Innovative studies have strived to provide new tools and materials for the development of systems offering new possibilities for industrial applications of enzymes. Herein, an effort has been made to present up-to-date developments on enzyme immobilization and current challenges in the food and beverage industries in terms of enhancing the enzyme stability.

Hepatocyte growth factor promotes the proliferation of human embryonic stem cell derived retinal pigment epithelial cells.

Research that pertains to the molecular mechanisms involved in retinal pigment epithelial (RPE) development can significantly contribute to cell therapy studies. The effects of periocular mesenchymal cells on the expansion of RPE cells remain elusive. We have examined the possible proliferative role of hepatocyte growth factor (HGF) as a mesenchymal cell secretory factor against human embryonic stem cell derived RPE (hESC-RPE). We found that the conditioned medium of human mesenchymal stem cells from apical papilla and/or exogenous HGF promoted proliferation of the hESC-RPE cells as single cells and cell sheets, in addition to rabbit RPE sheets in vitro. Blockage of HGF signaling by HGF receptor inhibitor, PHA-665752, inhibited proliferation of hESC-RPE cells. However, differentiation of hESCs and human-induced pluripotent stem cells to a rostral fate and eye-field specification was unaffected by HGF. Our in vivo analysis showed HGF expression in periocular mesenchymal cells after optic cup formation in chicken embryos. Administration of HGF receptor inhibitor at this developmental stage in chicken embryos led to reduced eye size and disorganization of the RPE sheet. These findings suggested that HGF administration could be beneficial for obtaining higher numbers of hESC-RPE cells in human preclinical and clinical trials.

Hedgehog signalling is dispensable in the proliferation of stem cells from human exfoliated deciduous teeth.

The hedgehog (Hh) signalling pathway is one of the key regulators in development with a dual role in cell fate specification, proliferation, and survival on different target cells. We have investigated the effect of recombinant sonic hedgehog (r-SHH) on extracted multipotent stem cells from human exfoliated deciduous teeth (SHED), which represent a potential stem cell population for therapeutic applications. Cell proliferation and cycle assays shown that r-SHH did not have a distinctive effect on cell cycle progression, nor did it increase cell number over a wide range of concentrations. Quantitative polymerase chain reaction (Q-PCR) also suggests that r-SHH treatment has no demonstrable influence on expression of proliferative genes (CCNE1 and KI67); in contrast, the anti-proliferative gene (CDKN1A) is overexpressed in response to SHH. Our findings have suggested the possibility that SHEDs demonstrate a different potential from human bone marrow mesenchymal stem cells (h-BMSCs) and dorsal neural progenitor in response to growth factors such as SHH.

Acetazolamide triggers death inducing autophagy in T-47D breast cancer cells.

The inhibitory effects of acetazolamide on the growth and proliferation of epithelial breast cancer cells (T-47D) were investigated. Analysis of morphological changes indicated little apoptosis in T-47D cells incubated with acetazolamide, according to data from flow cytometry, DNA laddering, and expression of AIF. However, an increase in caspase-3 activity was detected in cells. This was concomitant with an increase in DFF45/DFF40 ratio leading to inhibition of caspase-3 activity, DNA fragmentation and progression of apoptosis. Flow cytometry also confirmed that acetazolamide had no significant effect on cell cycle progression. These results are consistent with lack of change in the expression of cell cycle regulatory proteins p21, p27, cdc2 and cyclinD1. Increased expression of ATG5, p53 and DRAM, along with an increase in BCLN1/Bcl-2 ratio, indicated that acetazolamide inhibited the proliferation of T-47D cells by inducing autophagy. Increased expression of PTEN, along with decreased expression of Akt1, also showed that acetazolamide treatment resulted in death inducing autophagy. Collectively the results indicate that autophagy is an adequate mechanism mediating the anti-cancer effects of acetazolamide in T-47D cells through engagement of p53/DRAM pathway and attenuation of Akt survival signalling.

Biotechnological advancements towards water, food and medical healthcare: A review.

The global health status is highly affected by the growing pace of urbanization, new lifestyles, climate changes, and resource exploitation. Modern technologies pave a promising way to deal with severe concerns toward sustainable development. Herein, we provided a comprehensive review of some popular biotechnological advancements regarding the progress achieved in water, food, and medicine, as the most substantial fields related to public health. The emergence of novel organic/inorganic materials has brought about significant improvement in conventional water treatment techniques, anti-fouling approaches, anti-microbial agents, food processing, biosensors, drug delivery systems, and implants. Particularly, a growing interest has been devoted to nanomaterials and their application for developing novel structures or improving the characteristics of standard components. Also, bioinspired materials have been widely used to improve the performance, efficiency, accuracy, stability, safety, and cost-effectiveness of traditional systems. On the other side, the fabrication of innovative devices for precisely monitoring and managing various ecosystem and human health issues is of great importance. Above all, exceptional advancements in designing ion-selective electrodes (ISEs), microelectromechanical systems (MEMs), and implantable medical devices have altered the future landscape of environmental and biomedical research. This review paper aimed to shed light on the wide-ranging materials and devices that have been developed for health applications and mainly focused on the impact of nanotechnology in this field.

Isolation and characterization of human periodontal ligament stem cells under the terms of use in clinical application: A pilot study.

Background: The aim of the present study is to determine the possibility of isolation and characterization of the human periodontal ligament stem cells (hPDLSCs) using limited harvested periodontal ligament (PDL) tissue of only one patient's wisdom teeth (2-4 teeth) under the more compatible terms of use in clinical application without using the fetal bovine serum (FBS). Materials and Methods: In this pilot study, hPDLSCs were isolated from the impacted third molar, and tissue was scraped from the roots of the impacted third molar of 10 volunteers to enzymatically digest using collagenase. The cells were sub-cultured. The samples of the first seven patients and half of the eighth patient's sample were cultured in alpha modified of Eagle's medium (α-MEM) (-FBS) medium and the other part of the eighth patient's sample was cultured with prior medium supplemented with +FBS 15% as a control of the cultivation protocol. While for the past two patients (9th and 10th the α-MEM medium was supplemented with L-Glutamine, anti/anti 2X, and 20% knock-out serum replacement (KSR). Two more nutritious supplements (N2 and B27) were added to the medium of the tenth sample. Flow-cytometric analysis for the mesenchymal stem cell surface markers CD105, CD45, CD90, and CD73 was performed. Subsequent polymerase chain reaction was undertaken on three samples cultured with two growth media. Results: Cultivation failed in some of the samples because of the lack of cell adhesion to the culturing dish bottom (floating cells), but it was successful for the 9th and 10th patients, which were cultured in the α-MEM serum supplemented with KSR 20%. Flow cytometry analysis was positive for CD105, CD90, and CD73 and negative for CD45. The PDL stem cells (PDLSCs) expressed CD105, CD45, and CD90 but were poor for CD73. Conclusion: According to the limited number of sample tests in this study, isolation and characterization of PDLSCs from collected PDL tissue of one patient's wisdom teeth (2-4) may be possible by the proper setup in synthetic FBS-free serum.

Pumpless deterministic lateral displacement separation using a paper capillary wick.

Deterministic lateral displacement (DLD) is a passive separation method that separates particles by hydrodynamic size. This label-free method is a promising technique for cell separation because of its high size resolution and insensitivity to flow rate. Development of capillary-driven microfluidic technologies allows microfluidic devices to be operated without any external power for fluid pumping, lowering their total cost and complexity. Herein, we develop and test a DLD-based particle and cell sorting method that is driven entirely by capillary pressure. We show microchip self-filling, flow focusing, flow stability, and capture of separated particles. We achieve separation efficiency of 92% for particle-particle separation and more than 99% efficiency for cell-particle separation. The high performance of driven flow and separation along with simplicity of the operation and setup make it a valuable candidate for point-of-care devices.

Comparison of Adhesion and Proliferation of Human Gingival Fibroblasts on Acellular Dermal Matrix with and without Low Level Diode Laser Irradiation, an in vitro Study.

Statement of the Problem: In recent years, regeneration of periodontal soft tissues in the reconstruction of periodontal defects and the finding of suitable membranes and graft materials for the placement of autogenous grafts have been of great interest in various studies. In this regard, the proliferation and adhesion of regenerative cells are two linchpins of the complete regenerative process. Purpose: This study aimed to evaluate the effects of low-level laser beams on the attachment and the proliferation of human gingival fibroblasts in the presence of acellular dermal matrix (ADM). Materials and Method: All the experiments were conducted compared to tissue culture plate in four groups as follows: (1) Fibroblast+ADM+laser, (2) Fibroblast+ADM+ no laser, (3) Fibroblast + laser radiation, and (4) Fibroblast+ no laser. In this experimental study, the primary attachment was evaluated by passing 8h from seeding of 5×105 gingival fibroblasts with or without a single dose (15.6 J/cm2) of laser radiation. Cell proliferation rate was also examined at 24, 48, and 72 hours after cell culture, following exposure to 5.2 J/cm2 of laser at each day of examination. Thereafter, fibroblasts were incubated under the normal culture condition (at 37°C, 5% CO2) in high glucose Dulbecco's Modified Eagle's medium (DMEM) medium supplemented with 10% fetal bovine serum, 1% glutamax, and 1% penicillin/streptomycin. Subsequently, the cellular viability was assessed on each time point using MTS calorimetric assay. The obtained data were statistically analyzed by applying ANOVA and Tukey tests. Results: There was a significant difference among the means of these four groups in terms of the proliferation of fibroblasts at 24, 48 and 72 hours (p< 0.001). Moreover, there was no significant difference among the means of two groups in terms of fibroblastic attachment in 8 hours (p< 0.2). The fibroblast group has shown the highest proliferation rate among all groups after laser radiation. Conclusion: It was indicated that the laser radiation increases the fibroblast cell proliferation. Accordingly, although this increase was higher in the fibroblast group alone compared to the fibroblasts cultured on acellular dermal matrix, the laser radiation did not significantly increase the attachment of fibroblast cells to acellular dermal matrix.

Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists.

OBJECTIVE: Tendon repair strategies usually are accompanied by pathological mineralization and scar tissue formation that increases the risk of re-injuries. This study aimed to establish an efficient tendon regeneration method simultaneously with a reduced risk of ectopic bone formation. MATERIALS AND METHODS: In this experimental study, tenogenic differentiation was induced through transforming growth factor- ß3 (TGFB3) treatment in combination with the inhibiting concentrations of bone morphogenetic proteins (BMP) antagonists, gremlin-2 (GREM2), and a Wnt inhibitor, namely sclerostin (SOST). The procedure's efficacy was evaluated using real-time polymerase chain reaction (qPCR) for expression analysis of tenogenic markers and osteochondrogenic marker genes. The expression level of two tenogenic markers, SCX and MKX, was also evaluated by immunocytochemistry. Sirius Red staining was performed to examine the amounts of collagen fibers. Moreover, to investigate the impact of the substrate on tenogenic differentiation, the nanofibrous scaffolds that highly resemble tendon extracellular matrix was employed. RESULTS: Aggregated features formed in spontaneous normal culture conditions followed by up-regulation of tenogenic and osteogenic marker genes, including SCX, MKX, COL1A1, RUNX2, and CTNNB1. TGFB3 treatment exaggerated morphological changes and markedly amplified tenogenic differentiation in a shorter period of time. Along with TGFB3 treatment, inhibition of BMPs by GREM2 and SOST delayed migratory events to some extent and dramatically reduced osteo-chondrogenic markers synergistically. Nanofibrous scaffolds increased tenogenic markers while declining the expression of osteo-chondrogenic genes. CONCLUSION: These findings revealed an appropriate in vitro potential of spontaneous tenogenic differentiation of eq- ASCs that can be improved by simultaneous activation of TGFB and inhibition of osteoinductive signaling pathways.

Effect of process parameters on separation efficiency in a deterministic lateral displacement device.

Deterministic lateral displacement (DLD) is a hydrodynamic method known for its high-resolution sorting of particles. It achieves this through a periodic array of obstacles and laminar flow that passively directs particles along in two different directions depending on the particles' diameter. Many prior publications have been dedicated to the structural and geometrical development of DLD arrays to improve separation performance; however, a successful separation requires much more than a well-designed array. This paper shows how separation performance is affected by process parameters. For this purpose, the design and fabrication of a DLD device are described. Then three experiments show how process parameters affect the performance of the device. The first experiment uses dye solutions to visualize the formation of a hydrodynamically focused sample stream. The second experiment shows that the particle separation performance (of 7- & 15-µm particles) is affected by the way output fluids are collected. Finally, the third experiment looks at the particle separation efficiency as the input flow rates and the ratio of buffer to sample are changed. The results show that the proper range for buffer and sample flow rate in this device is 1-10 and 0.1-1 (µl/min), respectively. The buffer to sample flow rate ratio of 10 gives the highest separation efficiency, but at a lower sample throughput. The optimized values are specific for our device but demonstrate processes that we believe are universal for DLD separations.

In Vitro Study of the Recruitment and Expansion of Mesenchymal Stem Cells at the Interface of a Cu-Doped PCL-Bioglass Scaffold.

Developing new barrier membranes with improved biomechanical characteristics has acquired much interest owing to their crucial role in the field of periodontal tissue regeneration. In this regard, we enriched the electrospun polycaprolactone (PCL)/gelatin (Gel) membranes by adding bioglass (BG) or Cu-doped bioglass (CuBG) and examined their cellular adhesion and proliferation potential in the presence of alveolar bone marrow-derived mesenchymal stem cells (aBMSCs). The membranes were fabricated and characterized using mechanical strength, SEM, FTIR, EDX, and ICP assay. Besides, aBMSCs were isolated, characterized, and seeded with a density of 35,000 cells in each experimental group. Next, the cellular morphology, cell adhesion capacity, proliferation rate, and membrane antibacterial activity were assessed. The results displayed a significant improvement in the wettability, pore size, and Young's modulus of the PCL membrane following the incorporation of gelatin and CuBG particles. Moreover, all scaffolds exhibited reasonable biocompatibility and bioactivity in physiological conditions. Although the PCL/Gel/CuBG membrane revealed the lowest primary cell attachment, cells were grown properly and reached the confluent state after seven days. In conclusion, we found a reasonable level of attachment and proliferation of aBMSCs on all modified membranes. Meanwhile, a trace amount of Cu provided superiority for PCL/Gel/CuBG in periodontal tissue regeneration.

Assessment of Cytotoxicity and Odontogenic/Osteogenic Differentiation Potential of Nano-Dentine Cement Against Stem Cells from Apical Papilla.

OBJECTIVE: Assessment of the cytotoxicity of novel calcium silicate-based cement is imperative in endodontics. This experimental study aimed to assess the cytotoxicity and odontogenic/osteogenic differentiation potential of a new calcium silicate/pectin cement called Nano-dentine against stem cells from the apical papilla (SCAPs). MATERIALS AND METHODS: In this experimental study, the cement powder was synthesized by the sol-gel technique. Zirconium oxide was added as opacifier and Pectin, a plant-based polymer, and calcium chloride as the liquid to prepare the nano-based dental cement. Thirty-six root canal dentin blocks of human extracted single-canal premolars with 2 mm height, flared with #1, 2 and 3 Gates-Glidden drills were used to prepare the cement specimens. The cement, namely mineral trioxide aggregate (MTA), Biodentine, and the Nano-dentine were mixed according to the manufacturers' instructions and applied to the roots of canal dentin blocks. The cytotoxicity and odontogenic/osteogenic potential of the cement were evaluated by using SCAPs. RESULTS: SCAPs were characterized by the expression of routine mesenchymal cell markers and differentiation potential to adipocytes, osteoblasts, and chondrocytes. Cement displayed no significant differences in cytotoxicity or calcified nodules formation. Gene expression analysis showed that all three types of cement induced significant down- regulation of COLA1; however, the new cement induced significant up-regulation of RUNX2 and SPP1 compared to the control group and MTA. The new cement also induced significant up-regulation of TGFB1 and inducible nitric oxide synthase (iNOS) compared with Biodentine and MTA. CONCLUSION: The new Nano-dentin cement has higher odontogenic/osteogenic potential compared to Biodentine and MTA for differentiation of SCAPs to adipocytes, osteoblasts, and chondrocytes.

Cellular Behaviors of Periodontal Ligament Stem Cells in the Presence of Bone Grafting Biomaterials, In-Vitro Study.

Periodontal regeneration through the employment of bone substitutes has become a feasible strategy in animal and clinical studies. In this regard, we aimed to compare the periodontal ligament stem cell behavior in the vicinity of various bone grafting substitutes. Three types of popular bone substitutes, including allografts (Regen), xenografts (Cerabone), and alloplasts (Osteon) were studied in this experimental survey. The cellular attachment was assessed after four hours using the MTS assay and SEM imaging. In addition, cellular proliferation was investigated after 1, 3, 5, and 7 days through MTS assay. Osteogenesis was studied after 21 days of cell culture in a differentiation medium (DM+) and a normal medium (DM-), by employing real-time PCR and alizarin red staining. The highest cellular attachment was seen in the xenograft group with a significant difference in comparison to the other grafting materials. Despite the relatively low primary attachment of cells to allografts, the allograft group showed the highest total proliferation rate, while the lowest proliferation capacity was found in the alloplast group. Osteogenesis fount to be accelerated mostly by xenografts in both mediums (DM+ and DM-) after 3 weeks, while alloplasts showed the lowest osteogenesis. This study revealed that the type of bone substitutes used in regenerative treatments can affect cellular behavior and as a whole allografts and xenografts showed better results.

Recent advances in polymeric nanostructured ion selective membranes for biomedical applications.

Nano structured ion-selective membranes (ISMs) are very attractive materials for a wide range of sensing and ion separation applications. The present review focuses on the design principles of various ISMs; nanostructured and ionophore/ion acceptor doped ISMs, and their use in biomedical engineering. Applications of ISMs in the biomedical field have been well-known for more than half a century in potentiometric analysis of biological fluids and pharmaceutical products. However, the emergence of nanotechnology and sophisticated sensing methods assisted in miniaturising ion-selective electrodes to needle-like sensors that can be designed in the form of implantable or wearable devices (smartwatch, tattoo, sweatband, fabric patch) for health monitoring. This article provides a critical review of recent advances in miniaturization, sensing and construction of new devices over last decade (2011-2021). The designing of tunable ISM with biomimetic artificial ion channels offered intensive opportunities and innovative clinical analysis applications, including precise biosensing, controlled drug delivery and early disease diagnosis. This paper will also address the future perspective on potential applications and challenges in the widespread use of ISM for clinical use. Finally, this review details some recommendations and future directions to improve the accuracy and robustness of ISMs for biomedical applications.

Mechanobiology of Dental Pulp Stem Cells at the Interface of Aqueous-Based Fabricated ZIF8 Thin Film.

The limited knowledge on how biological systems sense and respond to the mechanical properties of metal-organic framework (MOF) thin films is a critical restriction factor for their extensive usage. To bridge this gap, we performed an in vitro study for defining and linking surface characteristics at the interface of the zeolitic imidazolate framework-8 (ZIF8) thin layer to stem cell behavior. First, the physio-mechanical properties of the ZIF8 layer grown on polydopamine (PDA) and tannic acid (TA) layers have been studied. The response of dental pulp stem cells (DPSCs) to different surface states was examined. The results showed that the uniform crystalline microstructure of the ZIF8 on PDA and TA effectively led to the 61- and 388-fold increased surface roughness, 3- and 2.5-fold moderated elastic modulus, almost 3-fold elevated surface free energy, and highly charged surfaces (ζ = -60 mV for TA/ZIF8), respectively. Beyond the inherent bioactivity of the ZIF8 layer, these substrate cues promoted advanced cell adhesion (∼two times) and high proliferation rate. Furthermore, we found a substantial increment in the differentiation efficiency of DPSCs on the ZIF8 layer, in a way that the expression of functional adipocyte (PPARG) and osteoblast (SPP1) markers was, respectively, elevated around 30 000- and 10 000-fold on the TA/ZIF8-coated silicon wafer (SW). Our findings support the impact of fabrication strategy on the biointerface properties of the ZIF8 layer and bring SW/TA/ZIF8 as a robust platform for managing stem cells for biomedical applications.