Primjena umjetne toplinske stimulacije kao novi pristup za indukciju tendinopatije u magaraca

Animal models for tendonitis are essential for studying the disease’s mechanism and pathogenesis, and evaluating different therapeutic protocols. The temperature seems to play a significant role in tendinitis initiation. The aim of this study was the generation of a novel, safe and cheap tendinitis model, and validation of its reliability. The effect of microwave diathermy (30 watts for 30 min) on the flexor tendons of donkeys as animal models was investigated after 15, 30 and 60 days. The evaluation was based on geometric analysis, ultrasonography, histomorphometric analysis and scanning electron microscopy (SEM). Microwave diathermy was capable of successfully inducing well-defined lesions in the superficial digital flexor tendon (SDFT) as well as the deep digital flexor tendon (DDFT). The results showed that all the animals exhibited signs of lameness, starting on day 15 and reaching on maximum on day 30. A significant increase in limb circumference was also detected on day 30 (P<0.05). Furthermore, the geometrical analysis of the proportion of induced lesion (PIL) in correlation with the tendon diameter, revealed that PIL was at the maximum width on day 30 (20.6 ± 1.2% for SDFT and 15.7 ± 0.7% for DDFT), as detected by ultrasound. Moreover, a high number of rounded tenocytes, bleeding, severe matrix disruption, and an increase in fiber thickness were detected by histomorphometric analysis. Also, the matrix alignment was severely disrupted in both SDFT and DDFT by day 30, as confirmed by SEM. In conclusion, using microwave diathermy for induction of tendonitis in donkey is a reliable, minimally invasive, and cost-effective tendonitis model.Uporaba životinjskih modela je neophodna za proučavanje mehanizma bolesti, patogeneze i za procjenu protokola liječenje tendinitisa. Čini se da temperatura igra značajnu ulogu pri nastajanju tendinitisa. Cilj istraživanja bio je stvaranje novog, sigurnog i jeftinog modela za proučavanje tendinitisa te potvrda njegove pouzdanosti. Učinak mikrovalne dijatermije (30 vata tijekom 30 min) na tetive fleksora u magaraca kao životinjskih modela istraživan je nakon 15, 30 i 60 dana. Procjena se temeljila na geometrijskoj analizi, ultrazvuku, histomorfometrijskoj analizi i skenirajućoj elektronskoj mikroskopiji (SEM). Mikrovalna dijatermija uspješno je inducirala dobro definirane lezije u površinskoj digitalnoj tetivi fleksora (SDFT) kao i dubokoj digitalnoj tetivi fleksora (DDFT). Rezultati su pokazali da su sve životinje pokazivale znakove hromosti, počevši od 15. dana i dostižući maksimum 30. dana. Također, 30. dana je uočeno znakovito (P<0,05) povećanje opsega ekstremiteta. Nadalje, uporabom ultrazvuka i primjenom geometrijske analize udjela inducirane lezije (PIL) u korelaciji s promjerom tetive, ustanovljeno je da je PIL bio maksimalne širine 30. dana (20,6 ± 1,2% za SFT i 15,7 ± 0,7% za DDFT). Osim toga, histomorfometrijskom analizom otkriven je povećan broj zaobljenih tenocita, krvarenje, teži poremećaji u matriksu i povećanje debljine vlakana. Do 30. dana, SEM analiza je pokazala izražene poremećaje u poravnavanju matriksa i u SDFT i u DDFT. Zaključno, primjena mikrovalne dijatermije za indukciju tendinitisa kod magaraca je pouzdan, minimalno invazivan i troškovno učinkovit životinjski model za proučavanje ove bolesti

A Hyaluronic Acid Demilune Scaffold and Polypyrrole-Coated Fibers Carrying Embedded Human Neural Precursor Cells and Curcumin for Surface Capping of Spinal Cord Injuries

[EN] Tissue engineering, including cell transplantation and the application of biomaterials and bioactive molecules, represents a promising approach for regeneration following spinal cord injury (SCI). We designed a combinatorial tissue-engineered approach for the minimally invasive treatment of SCI¿a hyaluronic acid (HA)-based scaffold containing polypyrrole-coated fibers (PPY) combined with the RAD16-I self-assembling peptide hydrogel (Corning® PuraMatrix¿peptide hydrogel (PM)), human induced neural progenitor cells (iNPCs), and a nanoconjugated form of curcumin (CURC). In vitro cultures demonstrated that PM preserves iNPC viability and the addition of CURC reduces apoptosis and enhances the outgrowth of Nestin-positive neurites from iNPCs, compared to nonembedded iNPCs. The treatment of spinal cord organotypic cultures also demonstrated that CURC enhances cell migration and prompts a neuron-like morphology of embedded iNPCs implanted over the tissue slices. Following sub-acute SCI by traumatic contusion in rats, the implantation of PMembedded iNPCs and CURC with PPY fibers supported a significant increase in neuro-preservation (as measured by greater III-tubulin staining of neuronal fibers) and decrease in the injured area (as measured by the lack of GFAP staining). This combination therapy also restricted platelet-derived growth factor expression, indicating a reduction in fibrotic pericyte invasion. Overall, these findings support PM-embedded iNPCs with CURC placed within an HA demilune scaffold containing PPY fibers as a minimally invasive combination-based alternative to cell transplantation alone.This research was funded by the Science by Women program, Women for Africa Foundation to H.E. and the grants FEDER/Ministerio de Ciencia e Innovacion-Agencia Estatal de Investigacion [RTI2018-095872-B-C21 and -C22/ERDF]; Part of the equipment employed in this work was funded by Generalitat Valenciana and cofinanced with ERDF funds (OP ERDF of Comunitat Valenciana 2014-2020). RISEUP project FetOpen in H2020 Program: H2020-FETOPEN-2018-2019-2020-01.Elkhenany, H.; Bonilla, P.; Giraldo-Reboloso, E.; Alastrue Agudo, A.; Edel, MJ.; Vicent, MJ.; Gisbert-Roca, F.... (2021). A Hyaluronic Acid Demilune Scaffold and Polypyrrole-Coated Fibers Carrying Embedded Human Neural Precursor Cells and Curcumin for Surface Capping of Spinal Cord Injuries. Biomedicines. 9(12):1-19. https://doi.org/10.3390/biomedicines9121928S11991

Green propolis extract promotes in vitro proliferation, differentiation, and migration of bone marrow stromal cells

Background Propolis is a resinous material extracted from bee glue with a complex chemical composition. The unique biological properties of propolis have led to its use in alternative medicine and as a nutritional supplement. Recent research shows that propolis could affect the immune system by decreasing the production of inflammatory cytokines and potentiating an effect on resident stem cells. The exact mechanism, however, is unknown. The goal of this study was to demonstrate whether green propolis extract affects any characteristic properties of mesenchymal stromal cells (MSCs)in vitro. Methods The cytocompatibility of propolis extract and the proliferation of bone marrow mesenchymal stromal cells (BMMSCs) in the presence of propolis was evaluated by live/dead cell staining and MTS viability assay over a period of 3 days. Also, we evaluated the effect of propolis extract on trilineage differentiation and migration capacity of undifferentiated and differentiated BMMSCs. Results Relative to the control, propolis extract resulted in a significant and linear increase in the proliferation of MSCs and inhibited the osteogenic differentiation of BMMSCs, while there was a potentiation of chondrogenesis and adipogenesis. Finally, in relevance to wound healing, an in vitro scratch assay demonstrated that the migratory potential of differentiated BMMSCs was enhanced in the presence of propolis. Conclusion We have demonstrated that propolis extract was not toxic to BMMSCs (\u3c400 \u3eμg/ml), supported their proliferation, potentiated chondrogenic and adipogenic differentiation processes, and supported cell migrationin vitro. Most interestingly, there was a down-regulation of osteogenesis. These data support the use of propolis extract for enhanced cell proliferation and tissue regeneration; however, it warrants further investigation

Ternary nanocomposite potentiates the lysophosphatidic acid effect on human MG63 osteoblast maturation

Aim: This study aimed to investigate the potential of ternary nanocomposite (TNC) to support MG63 osteoblast maturation to EB1089-(3S)1-fluoro-3-hydroxy-4-(oleoyloxy)butyl-1-phosphonate (FHBP) cotreatment. Materials & methods: Binary (P25/reduced graphene oxide [rGO]) nanocomposite was prepared, and silver (Ag) nanoparticles were loaded onto the surface to form TNC (P25/rGO/Ag). The influence of TNC on proliferation, alkaline phosphatase activity and osteogenic gene expression was evaluated in a model of osteoblast maturation wherein MG63 were costimulated with EB1089 and FHBP. Results: TNC had no cytotoxic effect on MG63. The addition of TNC to EB1089-FHBP cotreatment enhanced the maturation of MG63, as supported by the greater alkaline phosphatase activity and OPN and OCN gene expression. Conclusion: TNC could serve as a promising carrier for FHBP, opening up possibilities for its application in bone regeneration

Comparison of MXene and SWCNTs effectiveness for osteoblast differentiation: A preliminary study with MG-63 cells

The exceptional chemical and physical properties of nanomaterials have attracted growing interest in the field of bone tissue engineering. Single walled carbon nanotubes (SWCNTs) and MXenes are two types of unique nanomaterials that have shown promising potential for tissue engineering applications. In this study, we aimed to compare the differentiation capacity of MG-63 cells in the presence of carbon nanotubes (CNTs) and MXene nanomaterials. Methods: The cytotoxic effect of MXene and SWCNTs based nanocomposites was evaluated using the MTS assay. Also, the differentiation potential was evaluated using two models of stimulation. The first model was a co-stimulation of MG-63 s with EB1089 and an analogue of lysophosphatidic acid (LPA), (3S)-1-fluoro-3-hydroxy-4-(oleoyloxy)butyl-1-phosphonate (FHBP). The second model utilized β-glycerophosphate (βGP) and ascorbic acid (ASC). Results: MXene and SWCNTs showed no cytotoxic effect after 48 h of culture. MXene could enhance the maturation of MG-63 in both models of differentiation compared to controls and SWCNTs. Conclusion: The incorporation of MXene into MG-63 cell culture suggests that it has a promising potential as a biomaterial for bone regeneration as it demonstrated improved osteoblast maturation compared to SWCNTs

Ternary nanocomposite potentiates the lysophosphatidic acid effect on human osteoblast (MG63) maturation - supplementary data

Aim: This study aimed to investigate the potential of ternary nanocomposite (TNC) to support MG63 osteoblast maturation to EB1089-(3S)1-fluoro-3-hydroxy-4-(oleoyloxy)butyl-1-phosphonate (FHBP) cotreatment. Materials & methods: Binary (P25/reduced graphene oxide [rGO]) nanocomposite was prepared, and silver (Ag) nanoparticles were loaded onto the surface to form TNC (P25/rGO/Ag). The influence of TNC on proliferation, alkaline phosphatase activity and osteogenic gene expression was evaluated in a model of osteoblast maturation wherein MG63 were costimulated with EB1089 and FHBP. Results: TNC had no cytotoxic effect onMG63. The addition of TNC to EB1089-FHBP cotreatment enhanced the maturation of MG63, as supported by the greater alkaline phosphatase activity and OPN and OCN gene expression. Conclusion: TNC could serve as a promising carrier for FHBP, opening up possibilities for its application in bone regeneration.Plain language summary: Nanoparticles (NPs) are often used in medicine because they have certain benefits over traditional drugs, such as increased delivery. Multiple NPs can be combined into hybrid NPs called nanocomplexes, which can have many positive effects. One application of nanomedicine is to encourage the repair of certain body tissues such as bones. Encouraging stem cells to differentiate into bone cells and immature bone cells to mature is key in this process. This study made a ternary nanocomplex (TNC), meaning it was comprised of three NPs. This TNC was designed to deliver a drug called (3S)1-fluoro- 3-hydroxy-4-(oleoyloxy)butyl-1-phosphonate (FHBP), which has been shown to encourage the maturation and development of osteoblasts, a type of bone cell. The TNC was made up of silver NPs, which can kill bacteria; reduced graphene oxide, which enhances the production of bone cells; and titanium dioxide, which has shown effectiveness in wound healing and mixed results in bone tissue regeneration. This TNC was tested on a cell line that comes from a type of bone cancer called MG63. The TNC was found to not be toxic to these cells. TNC incorporation into FHBP treatment enhanced the maturation of MG63. This suggests that these TNCs could be an effective treatment to encourage bone repair following joint replacement surgeries.</p

Curcumin-infused nanostructured lipid carriers: a promising strategy for enhancing skin regeneration and combating microbial infection

Abstract Background Curcumin is a biomolecule that can be extracted from the Curcuma longa that has been shown to have the potential to aid skin wound healing. It has been studied for its anti-inflammatory and antioxidant properties, which may help to reduce swelling and promote tissue repair. However, curcumin has low solubility in water, which can limit its absorption and bioavailability. Encapsulating it in lipid nanoparticles may help to increase its absorption, leading to improved bioavailability. Methods Curcumin-loaded nanostructure lipid nanocarriers (CURC-NLCs) were prepared and characterized. Also, the phenolic, flavonoid contents, antioxidant and antimicrobial efficacy against gram-positive and gram-negative bacteria were investigated. Furthermore, in vivo rabbit animal model was used to test its regenerative capacity and wound-healing efficiency. Results The CURC-NLCs significantly increased the content of phenolic and flavonoid compounds compared to curcumin, resulting in a dramatic increase in antioxidant activity. CURC-NLCs also showed a potent inhibitory effect on Gram-positive, Gram-negative, and fungi, two times higher than curcumin. CURC-NLCs showed a higher potential to fasten the wound healing of full-thickness skin injuries as it resulted in 1.15- and 1.9-fold higher wound closure at the first week of injury compared to curcumin and control, respectively (p < 0.0001). Conclusion These results suggest that CURC-NLCs have an excellent potential to promote skin regeneration, which could be attributed to its antioxidant and broad-spectrum antimicrobial effect

Telomerase reverse transcriptase coordinates with the epithelial-to-mesenchymal transition through a feedback loop to define properties of breast cancer stem cells

Telomerase and its core component, telomerase reverse transcriptase (hTERT), are critical for stem cell compartment integrity. Normal adult stem cells have the longest telomeres in a given tissue, a property mediated by high hTERT expression and high telomerase enzymatic activity. In contrast, cancer stem cells (CSCs) have short telomeres despite high expression of hTERT, indicating that the role of hTERT in CSCs is not limited to telomere elongation and/or maintenance. The function of hTERT in CSCs remains poorly understood. Here, we knocked down hTERT expression in CSCs and observed a morphological shift to a more epithelial phenotype, suggesting a role for hTERT in the epithelial-to-mesenchymal transition (EMT) of CSCs. Therefore, in this study, we systematically explored the relationship between hTERT and EMT and identified a reciprocal, bi-directional feedback loop between hTERT and EMT in CSCs. We found that hTERT expression is mutually exclusive to the mesenchymal phenotype and that, reciprocally, loss of the mesenchymal phenotype represses hTERT expression. We also showed that hTERT plays a critical role in the expression of key CSC markers and nuclear β-catenin localization, increases the percentage of cells with side-population properties, and upregulates the CD133 expression. hTERT also promotes chemoresistance properties, tumorsphere formation and other important functional CSC properties. Subsequently, hTERT knockdown leads to the loss of the above advantages, indicating a loss of CSC properties. Our findings suggest that targeting hTERT might improve CSCs elimination by transitioning them from the aggressive mesenchymal state to a more steady epithelial state, thereby preventing cancer progression