Analysis of gene expression profiles of Enterococcus faecalis induced by type I collagen

Enterococcus faecalis is an etiological agent of endodontic infections. The present study was performed to investigate the gene profiles of E. faecalis induced by type I collagen stimulation. E. faecalis ATCC 19433 was cultivated with [collagen (+)] or without type I collagen [collagen (−)], and transcriptome analysis was performed using high-throughput sequencing technology. A total of 3.6 gb of information was obtained by sequence analysis and 77 differentially expressed genes (DEGs) between the two culture conditions were identified. Among the 77 DEGs, 35 genes were upregulated in collagen (+) E. faecalis, whereas 42 genes were downregulated. Gene Ontology (GO) enrichment analysis was performed and 11 GO terms, including metalloendopeptidase activity (GO:0004222) and two related GO terms (GO:0031012, GO:0044421), were significantly enriched in the set of upregulated genes. We focused on an upregulated DEG belonging to the matrixin metalloprotease gene family, and matrix metalloprotease (MMP) activities of the bacterial cell were examined. The generic MMP, MMP-8, and MMP-9 activities of collagen (+) E. faecalis were significantly higher than those of collagen (−) E. faecalis. These results suggested that contact with type I collagen may alter the gene expression profile of E. faecalis, and upregulation of metalloprotease genes may result in enhanced MMP activities in E. faecalis.

Energy Alterations in Patient with Osteogenesis Imperfecta

Introduction; Osteogenesis imperfecta is according to Western medicine, a disorder of the connective tissue caused by an abnormal synthesis or processing type I collagen of genetic origin, a protein that is important to strengthen bones. The clinical manifestation of this problem can cause blue sclera, short stature, and deafness in adulthood, dentinogenesis imperfecta. In traditional Chinese medicine (TCM), osteogenesis imperfecta is related to Kidney energy deficiency (second chakra). Purpose: the purpose of this study is to show that patients with osteogenesis imperfecta has energy deficiency in the Kidney energy (second chakra) and the treatment of this condition, replenishing this energy using highly diluted medications is very important to treat the root of the problem and not just treating the symptoms. Methods: through one case report of 30 years-old man with history of several fractures since childhood. He went acupuncture clinic to treat his anxiety symptoms and I saw that his sclera was blue. Treatment was done using Chinese dietary counseling, auricular acupuncture with apex ear bloodletting and systemic acupuncture. Radiesthesia procedure were used to measure his chakras’ energy centers. Results: All his chakras’ were in the lowest level of energy, including the second that was the Kidney, responsible for the bone and teeth. The treatment began replenishing this chakras’ energy centers using highly diluted medications, such as homeopathies, according to the theory created by me (2020) entitled Constitutional Homeopathy of the Five Elements Based on Traditional Chinese Medicine and crystal-based medications. Conclusion: through this case reported in this article, I can say that patient with osteogenesis imperfecta has energy deficiency in the five internal massive organ, especially in the Kidney and the treatment of these energy deficiency, is very important to treat patients with osteogenesis imperfecta in the deepest level, in the energy point of view.

Serum levels of C-terminal telopeptide (CTX) are associated with muscle function in community-dwelling older adults

Background Markers of bone metabolism have been associated with muscle mass and function. Whether serum cross-linked C-terminal telopeptides of type I collagen (CTX) is also associated with these measures in older adults remains unknown. Methods In community-dwelling older adults at high risk of falls and fractures, serum CTX (biochemical immunoassays) was used as the exposure, while appendicular lean mass (dual-energy x-ray absorptiometry) and muscle function (grip strength [hydraulic dynamometer], short physical performance battery [SPPB], gait speed, sit to stand, balance, Timed Up and Go [TUG]) were used as outcomes. Potential covariates including demographic, lifestyle and clinical factors were considered in statistical models. Areas under the ROC curves were calculated for significant outcomes. Results 299 older adults (median age: 79 years, IQR: 73, 84; 75.6% women) were included. In multivariable models, CTX was negatively associated with SPPB (β = 0.95, 95% CI: 0.92, 0.98) and balance (β = 0.92, 0.86, 0.99) scores, and positively associated with sit to stand (β = 1.02, 95% CI: 1.00, 1.05) and TUG (β = 1.03, 95% CI: 1.00, 1.05). Trend line for gait speed (β = 0.99, 95% CI: 0.98, 1.01) was in the hypothesized direction but did not reach significance. AUC curves showed low diagnostic power (<0.7) of CTX in identifying poor muscle function (SPPB: 0.63; sit to stand: 0.64; TUG: 0.61). Conclusion In older adults, higher CTX levels were associated with poorer lower-limb muscle function (but showed poor diagnostic power for these measures). These clinical data build on the biomedical link between bone and muscle.

Incorporation of Superparamagnetic Iron Oxide Nanoparticles into Collagen Formulation for 3D Electrospun Scaffolds

Nowadays, there is an ever-increasing interest in the development of systems able to guide and influence cell activities for bone regeneration. In this context, we have explored for the first time the combination of type-I collagen and superparamagnetic iron oxide nanoparticles (SPIONs) to design magnetic and biocompatible electrospun scaffolds. For this purpose, SPIONs with a size of 12 nm were obtained by thermal decomposition and transferred to an aqueous medium via ligand exchange with dimercaptosuccinic acid (DMSA). The SPIONs were subsequently incorporated into type-I collagen solutions to prove the processability of the resulting hybrid formulation by means of electrospinning. The optimized method led to the fabrication of nanostructured scaffolds composed of randomly oriented collagen fibers ranging between 100 and 200 nm, where SPIONs resulted distributed and embedded into the collagen fibers. The SPIONs-containing electrospun structures proved to preserve the magnetic properties of the nanoparticles alone, making these matrices excellent candidates to explore the magnetic stimuli for biomedical applications. Furthermore, the biological assessment of these collagen scaffolds confirmed high viability, adhesion, and proliferation of both pre-osteoblastic MC3T3-E1 cells and human bone marrow-derived mesenchymal stem cells (hBM-MSCs).

Evaluation of Polycaprolactone Electrospun Nanofiber-Composites for Artificial Skin Based on Dermal Fibroblast Culture

The study’s aim was to develop a dermal equivalent scaffold that can mimic the architecture and biological performance of the human dermis. Poly ε-caprolactone (PCL) electrospun nanofiber material (ENF) was assembled with polyethylene glycol diacrylate (PEGDA), sodium alginate (SA) and type I collagen (CG1) to develop three groups of dermal equivalent scaffolds. These scaffolds were named PEGDA-PCL, SA-PCL and CG1-PCL. Scanning electron microscopy (SEM) images of cell-free scaffolds’ top and cross-sectional surface were collected and analyzed to examine internal morphology, specifically the adhesiveness of PCL fibers with the different scaffolds. Human dermal fibroblasts were cultured on each of the scaffolds. Cell viability studies including cell adhesion, cell differentiation and stress fiber production were conducted on each scaffold. Furthermore, the architectural integrity of each scaffold was verified by degradation analysis for 2 weeks by soaking each scaffold in phosphate-buffered saline (PBS) solution. Finally, we conducted rheological characteristics of each scaffold. Based on our results from the above analysis, the study concluded that CG1-PCL is best suitable for the dermal equivalent model and has potential to be used as a graft for skin repair.

Investigating the histological and structural properties of tendon gel as an artificial biomaterial using the film model method in rabbits

Purpose This study aimed to evaluate the properties of tendon gel by investigating the histological and structural differences among tendon gels under different preservation periods using a rabbit model. Methods Forty mature female rabbits were divided into four groups, each containing ten rabbits, on the basis of in-vivo preservation periods of tendon gels (3, 5, 10, and 15 days). We created the Achilles tendon rupture models using the film model method to obtain tendon gels. Tensile stress was applied to the tendon gel to promote maturation. Histological and structural evaluations of the tendon gel were performed before and after applying the tensile force, and the results obtained from the four groups were compared. Results Although the day-3 and day-5 tendon gels before applying tensile stress were histologically more immature than the day-10 and day-15 gels, type I collagen fibers equivalent to those of normal tendons were observed in all groups after the tensile process. Based on the surface and molecular structural evaluations, the day-3 tendon gels after the tensile process were molecularly cross-linked, and thick collagen fibers similar to those present in normal tendons were observed. Structural maturation observed in the day-3 tendon gels caused by traction was hardly observed in the day-5, -10, and -15 tendon gels. Conclusions The day-3 tendon gel had the highest regenerative potential to become a normal tendon by applying a traction force.

Characteristics and prediction of subclinical hypocalcemia in dairy cows during the transition period using blood analytes

This study aimed to present the characteristics of and to predict subclinical hypocalcemia in dairy cows during the transition period using blood analytes. We examined fluctuations in plasma calcium (Ca), phosphorus (P), bone metabolic markers carboxy-terminal telopeptide of type I collagen (CTX), fibroblast growth factor (FGF23), 1,25(OH)2D3, parathyroid hormone, and other blood biochemical analytes from prepartum week 2 to postpartum day 14 in 116 multiparous high-producing Holstein cows from a free-stall barn dairy farm. With a plasma concentration of Ca <2.0 mmol/L as a criterion for the diagnosis of subclinical hypocalcemia, 64 cows were classified as normocalcemic, and 52 cows as subclinically hypocalcemic. Among the 52 hypocalcemic cows, 50 were detected on postpartum days 1 or 3, and 2 on postpartum day. The subclinically hypocalcemic cows were in a state of low bone turnover in the prepartum period, with low plasma concentrations of Ca and CTX. The subclinically hypocalcemic cows showed signs of a P regulation disorder in the prepartum period. This was marked by high plasma concentrations of P and low concentrations of 1,25(OH)2D3 and FGF23, which is also considered to be the cause of the low bone turnover. The results of a multiple logistic regression model showed that prepartum plasma concentrations of FGF23, CTX, and Ca were ideal predictors of postpartum subclinical hypocalcemia in dairy cows, using the model equation 38.8-0.052*FGF23-0.492*CTX-10.645*Ca, with a score of > 0 considered as an indication of increased risk of subclinical hypocalcemia after calving. The scoring rule had an accuracy of 79.3%, sensitivity of 76.9%, and specificity of 81.3%. The plasma concentrations of FGF23, CTX, and Ca were ideal predictors of postpartum subclinical hypocalcemia in dairy cows.

Model Systems for Evidencing the Mediator Role of Riboflavin in the UVA Cross-Linking Treatment of Keratoconus

Riboflavin under UVA radiation generates reactive oxygen species (ROS) that can induce various changes in biological systems. Under controlled conditions, these processes can be used in some treatments for ocular or dermal diseases. For instance, corneal cross-linking (CXL) treatment of keratoconus involves UVA irradiation combined with riboflavin aiming to induce the formation of new collagen fibrils in cornea. To reduce the damaging effect of ROS formed in the presence of riboflavin and UVA, the CXL treatment is performed with the addition of polysaccharides (dextran). Hyaluronic acid is a polysaccharide that can be found in the aqueous layer of the tear film. In many cases, keratoconus patients also present dry eye syndrome that can be reduced by the application of topical solutions containing hyaluronic acid. This study presents physico-chemical evidence on the effect of riboflavin on collagen fibril formation revealed by the following methods: differential scanning microcalorimetry, rheology, and STEM images. The collagen used was extracted from calf skin that contains type I collagen similar to that found in the eye. Spin trapping experiments on collagen/hyaluronic acid/riboflavin solutions evidenced the formation of ROS species by electron paramagnetic resonance measurements.

Modulation by 17,20S(OH)2pD of Fibrosis-Related Mediators in Dermal Fibroblast Lines from Healthy Donors and from Patients with Systemic Sclerosis

We previously demonstrated that the non-calcemic pregnacalciferol (pD) analog 17,20S (OH)2pD suppressed TGF-β1-induced type I collagen production in cultured normal human dermal fibroblasts. In the present studies, we examined fibroblasts cultured from the lesional skin of patients with systemic sclerosis (scleroderma (SSc)) and assessed the effects of 17,20S(OH)2pD on fibrosis-related mediators. Dermal fibroblast lines were established from skin biopsies from patients with SSc and healthy controls. Fibroblasts were cultured with either 17,20S(OH)2pD or 1,25(OH)2D3 (positive control) with/without TGF-β1 stimulation and extracted for protein and/or mRNA for collagen synthesis and mediators of fibrosis (MMP-1, TIMP-1, PAI-1, BMP-7, PGES, GLI1, and GLI2). 1 7,20S(OH)2pD (similar to 1,25(OH)2D3) significantly suppressed net total collagen production in TGF-β1-stimulated normal donor fibroblast cultures and in cultures of SSc dermal fibroblasts. 17,20S(OH)2pD (similar to 1,25(OH)2D3) also increased MMP-1, BMP-7, and PGES and decreased TIMP-1 and PAI1 expression in SSc fibroblasts. Although 17,20S(OH)2pD had no effect on Gli1 or Gli2 in SSc fibroblasts, it increased Gli2 expression when cultured with TGF-β1 in normal fibroblasts. These studies demonstrated that 17,20S(OH)2pD modulates mediators of fibrosis to favor the reduction of fibrosis and may offer new noncalcemic secosteroidal therapeutic approaches for treating SSc and fibrosis.

Assessment of Collagen-Based Nanostructured Biomimetic Systems with a Co-Culture of Human Bone-Derived Cells

Osteoporosis is a worldwide disease resulting in the increase of bone fragility and enhanced fracture risk in adults. In the context of osteoporotic fractures, bone tissue engineering (BTE), i.e., the use of bone substitutes combining biomaterials, cells, and other factors, is considered a potential alternative to conventional treatments. Innovative scaffolds need to be tested in in vitro systems where the simultaneous presence of osteoblasts (OBs) and osteoclasts (OCs), the two main players of bone remodeling, is required to mimic their crosstalk and molecular cooperation. To this aim, two composite materials were developed, based on type I collagen, and containing either strontium-enriched mesoporous bioactive glasses or rod-like hydroxyapatite nanoparticles. The developed nanostructured systems underwent genipin chemical crosslinking and were then tested with an indirect co-culture of human trabecular bone-derived OBs and buffy coat-derived OC precursors, for 2–3 weeks. The favorable structural and biological properties of the materials proved to successfully support the viability, adhesion, and differentiation of cells, encouraging a further investigation of the developed bioactive systems as biomaterial inks for the 3D printing of more complex scaffolds for BTE.