A randomized trial using telehealth technology to link caregivers with dementia care experts for in-home caregiving support: FamTechCare protocol.

The number of persons with dementia (PWD) in the United States is expected to reach 16 million by 2050. Due to the behavioral and psychological symptoms of dementia, caregivers face challenging in-home care situations that lead to a range of negative health outcomes such as anxiety and depression for the caregivers and nursing home placement for PWD. Supporting Family Caregivers with Technology for Dementia Home Care (FamTechCare) is a multisite randomized controlled trial evaluating the effects of a telehealth intervention on caregiver well-being and PWD behavioral symptoms. The FamTechCare intervention provides individualized dementia-care strategies to in-home caregivers based on video recordings that the caregiver creates of challenging care situations. A team of dementia care experts review videos submitted by caregivers and provide interventions to improve care weekly for the experimental group. Caregivers in the control group receive feedback for improving care based on a weekly phone call with the interventionist and receive feedback on their videos at the end of the 3-month study. Using linear mixed modeling, we will compare experimental and control group outcomes (PWD behavioral symptoms and caregiver burden) after 1 and 3 months. An exploratory descriptive design will identify a typology of interventions for telehealth support for in-home dementia caregivers. Finally, the cost for FamTechCare will be determined and examined in relation to hypothesized effects on PWD behavioral symptoms, placement rates, and caregiver burden. This research will provide the foundation for future research for telehealth interventions with this population, especially for families in rural or remote locations.

Nanoplex-Mediated Codelivery of Fibroblast Growth Factor and Bone Morphogenetic Protein Genes Promotes Osteogenesis in Human Adipocyte-Derived Mesenchymal Stem Cells.

This study highlights the importance of transfection mediated coordinated bone morphogenetic protein 2 (BMP-2) and fibroblast growth factor 2 (FGF-2) signaling in promoting osteogenesis. We employed plasmids independently encoding BMP-2 and FGF-2 complexed with polyethylenimine (PEI) to transfect human adipose derived mesenchymal stem cells (hADMSCs) in vitro. The nanoplexes were characterized for size, surface charge, in vitro cytotoxicity, and transfection ability in hADMSCs. A significant enhancement in BMP-2 protein secretion was observed on day 7 post-transfection of hADMSCs with PEI nanoplexes loaded with both pFGF-2 and pBMP-2 (PEI/(pFGF-2+pBMP-2)) versus transfection with PEI nanoplexes of either pFGF-2 alone or pBMP-2 alone. Osteogenic differentiation of transfected hADMSCs was determined by measuring osteocalcin and Runx-2 gene expression using real time polymerase chain reactions. A significant increase in the expression of Runx-2 and osteocalcin was observed on day 3 and day 7 post-transfection, respectively, by cells transfected with PEI/(pFGF-2+pBMP-2) compared to cells transfected with nanoplexes containing pFGF-2 or pBMP-2 alone. Alizarin Red staining and atomic absorption spectroscopy revealed elevated levels of calcium deposition in hADMSC cultures on day 14 and day 30 post-transfection with PEI/(pFGF-2+pBMP-2) compared to other treatments. We have shown that codelivery of pFGF-2 and pBMP-2 results in a significant enhancement in osteogenic protein synthesis, osteogenic marker expression, and subsequent mineralization. This research points to a new clinically translatable strategy for achieving efficient bone regeneration.

Analysis of the attachment and differentiation of three-dimensional rotary wall vessel cultured human preosteoblasts on dental implant surfaces.

PURPOSE: The purpose of this study was to determine whether osseous tissues engineered in three-dimensional (3D) environments preserved their mineralizing capacity and retained biologic characteristics when cultured on dental implant surfaces. MATERIALS AND METHODS: Human preosteoblast cells were cultured in both 3D rotary wall vessels and on 2D tissue culture plastic plates for 3 days. Aggregates from the 3D chambers and cells from the 2D plates were collected and transferred to commercially pure titanium disks with either 600-grit polished or sandblasted surfaces. These were cultured for an additional 7 days. The aggregates and cells from the disks were collected and prepared for scanning electron microscopy for microscopic evaluation and atomic adsorption assays for mineral content analysis. Additionally, staining with Alizarin red S was performed to compare the mineralization amount and pattern in each group. Polymerase chain reaction analysis was performed to evaluate expression of osteogenic genes, including Runx2, FAK, bone morphogenetic protein 2, and osteocalcin. RESULTS: Cells from 3D rotary wall vessel cultures attached to implant surfaces and presented cell attachment and growth patterns similar to those of standard 2D cultured cells, showing evidence of radial and random growth, yet they formed multiple focal niches on implant surfaces out of which cells proliferated. The 3D cultured cells and osseous tissues retained higher amounts of mineral formed during the initial culture and showed a higher tendency toward mineralization on implant surfaces compared to standard cultured cells. The 3D cultured cells and osseous tissues on implant surfaces at 1 week showed higher key gene protein expression. RNA expression at 1 week was equivalent to that of standard cultured cells. CONCLUSION: Culture of human osteogenic cells and tissues in 3D rotary wall vessels may expedite the osseointegration process on dental implant surfaces, thus reducing the overall treatment time.

Gene-Activated Titanium Surfaces Promote In Vitro Osteogenesis.

PURPOSE: Commercially pure titanium (CpTi) and its alloys possess favorable mechanical and biologic properties for use as implants in orthopedics and dentistry. However, failures in osseointegration still exist and are common in select individuals with risk factors such as smoking. Therefore, in this study, a proposal was made to enhance the potential for osseointegration of CpTi discs by coating their surfaces with nanoplexes comprising polyethylenimine (PEI) and plasmid DNA (pDNA) encoding bone morphogenetic protein-2 (pBMP-2). MATERIALS AND METHODS: The nanoplexes were characterized for size and surface charge with a range of N/P ratios (the molar ratio of amine groups of PEI to phosphate groups in pDNA backbone). CpTi discs were surface characterized for morphology and composition before and after nanoplex coating using scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and X-ray powder diffraction (XRD). The cytotoxicity and transfection ability of CpTi discs coated with nanoplexes of varying N/P ratios in human bone marrow-derived mesenchymal stem cells (BMSCs) was measured via MTS assays and flow cytometry, respectively. RESULTS: The CpTi discs coated with nanoplexes prepared at an N/P ratio of 10 (N/P-10) were considered optimal, resulting in 75% cell viability and 14% transfection efficiency. Enzyme-linked immunosorbent assay results demonstrated a significant enhancement in BMP-2 protein secretion by BMSCs 7 days posttreatment with PEI/pBMP-2 nanoplexes (N/P-10) compared to the controls, and real-time PCR data demonstrated that the BMSCs treated with PEI/pBMP-2 nanoplex-coated CpTi discs resulted in an enhancement of Runx-2, alkaline phosphatase, and osteocalcin gene expressions on day 7 posttreatment. In addition, these BMSCs demonstrated enhanced calcium deposition on day 30 posttreatment as determined by qualitative (alizarin red staining) and quantitative (atomic absorption spectroscopy) assays. CONCLUSION: It can be concluded that PEI/pBMP-2 nanoplex (N/P-10)-coated CpTi discs have the potential to induce osteogenesis and enhance osseointegration.

Effects of fluoride-modified titanium surfaces on osteoblast proliferation and gene expression.

PURPOSE: The objective of this study was to test the hypothesis that fluoride-modified titanium surfaces would enhance osteoblast differentiation. Osteoblast growth on a moderately rough etched fluoride-modified titanium surface (alteration in cellular differentiation) was compared to osteoblast growth on the same surface grit-blasted with titanium dioxide. The potential role of nanometer-level alterations on cell shape and subsequent differentiation was then compared. MATERIALS AND METHODS: Human embryonic palatal mesenchymal (HEPM) cultures were incubated on the respective surfaces for 1, 3, and 7 days, followed by analysis for cell proliferation, alkaline phosphatase (ALP) -specific activity, and mRNA steady-state expression for bone-related genes (ALP, type I collagen, osteocalcin, bone sialoprotein [BSP] II, Cbfa1, and osterix) by real-time polymerase chain reaction (PCR). RESULTS: The different surfaces did not alter the mRNA expression for ALP, type I collagen, osterix, osteocalcin, or BSP II. However, Cbfa1 expression on the fluoride-modified titanium surface was significantly higher (P < .001) at 1 week. The number of cells on this surface was 20% lower than the number of cells on the surface TiO2-blasted with 25-microm particles but not significantly different from the number of cells on the surface TiO2-blasted with 125-microm particles. Cells grown on all the titanium surfaces expressed similar levels of ALP activity. CONCLUSIONS: The results indicated that a fluoride-modified surface topography, in synergy with surface roughness, may have a greater influence on the level of expression of Cbfa1 (a key regulator for osteogenesis) than the unmodified titanium surfaces studied.

Differentiation of preosteoblasts is affected by implant surface microtopographies.

Osteogenesis involves the recruitment of multipotent mesenchymal cells and the progressive differentiation of these cells into osteoblasts. The transcription factor Runx2 regulates osteoblast differentiation and expression of genes necessary for the development of a mineralized phenotype. The purpose of this study was to determine if preosteoblast cell differentiation associated with Runx2 and osteocalcin gene expression was influenced by implant surface microtopography. Human embryonic palatal mesenchymal cells (HEPM cells) were cultured on grooved or roughened cpTi implant discs. Cell phenotypes were evaluated with epifluorescent microscopy. Real-time PCR was used for quantitative analysis of Runx2 and osteocalcin gene expression. HEPM cells mineralized when grown on rough and grooved implant surfaces relative to tissue culture plastic. Real-time PCR showed significant (p < 0.05) increases in Runx2 and osteocalcin gene expression in cells cultured on rough and grooved implant microtopographies. These results suggest that preosteoblast cell differentiation is affected by implant surface microtopographies during osseointegration of dental implants.

Integrin-associated tyrosine kinase FAK affects Cbfa1 expression.

Following cell adhesion, focal adhesion kinase (FAK) autophosphorylates on tyrosine and regulates intracellular signaling cascades that regulate cell growth and differentiation. The hypothesis of this study was FAK mediates osteoblast differentiation dependent Cbfa1 expression. Slowly mineralizing UI and rapidly mineralizing UMR-106-01 BSP osteoblasts formed focal adhesions; however, the level of FAK in UI focal adhesions was less than that seen in BSP cells. UI cultures had less FAK expression (p < 0.05) along with elevated levels of FAK phosphotyrosine in comparison to rapidly mineralizing BSP cultures. Mineralization decreased in a dose-dependent manner in response to Herbimycin A, a tyrosine kinase inhibitor. Overexpression of FAK in UI cells led to a fourfold increase in Cbfa1 gene expression (p < 0.02), and an increase in Cbfa1 protein expression. These results suggest that the integrin-associated tyrosine kinase FAK contributes to the regulation of the osteoblast differentiation in part through the regulation of Cbfa1 expression.

Intracellular release of 17-ß estradiol from cationic polyamidoamine dendrimer surface-modified poly (lactic-co-glycolic acid) microparticles improves osteogenic differentiation of human mesenchymal stromal cells.

Human bone marrow mesenchymal stromal cells (MSCs) are considered a potential cell source for MSC-based bone regeneration, but improvements in the proliferation and differentiation capacity of MSCs are necessary for practical applications. Estrogen effectively improves MSC capabilities and has strong potential as a regulator of MSCs. The aim of this study was to develop a delivery system that provides intracellular release of estrogen and test its ability to improve osteogenic differentiation of MSCs. Biodegradable poly (lactic-co-glycolic acid) (PLGA) microparticles were developed that entrap 17-ß estradiol (E2) and provide intracellular release of E2. The results show that we can prepare PLGA particles with efficient loading of E2 and maintain release of E2 up to 7 days. Surface modifying E2-loaded PLGA particles with cationic polyamidoamine dendrimers enabled increased uptake by human MSCs. Human MSC uptake of the E2-loaded PLGA particles significantly upregulates osteogenic differentiation markers of alkaline phosphatase and osteocalcin. In conclusion, cationic-modified PLGA particles can serve as a tool for intracellular delivery of estrogen to effectively execute estrogen regulation of MSCs. This approach has the potential to improve the osteogenic capabilities of MSCs and to develop appropriate environments of implantation for MSC-based bone tissue engineering.

Osteoblast differentiation is enhanced in rotary cell culture simulated microgravity environments.

PURPOSE: As the aging population increases, more people will become reliant on regenerative dental medicine for implant therapy. The objective of this study was to test the hypothesis that 3D rotary cell culture (RCC) environments created by simulated microgravity would enhance osteogenic gene expression using integrin mediated pathways. MATERIALS AND METHODS: Human embryonic palatal mesenchymal (HEPM, ATCC 1486) pre-osteoblasts were cultured in either RCC to create 3D environments or in 2D monolayers for 72 hours. Gross phenotypic analysis was performed using Alizarin Red S staining for calcium and microscopy. Real-time PCR analysis was used to detect differences in osteoblast gene expression. Aggregates developed in 3D RCC environments were treated with or without antibody to the collagen-I integrin receptor alpha2beta1 to determine whether this molecular pathway might contribute to the development of a mineralized matrix. RESULTS: Microscopic analysis demonstrated that RCC environments promoted 3D aggregate formation by 72 hours without any scaffold. The mass appeared osseous-like with a white, shiny, translucent surface. The center was amorphous with areas of vacuolization, tubule-like structures, and fibrous-like extensions. Real-time PCR data showed that 3D environments enhanced osteogenic gene expression as compared with 2D monolayer culturing conditions. At 72 hours, changes in levels of osteogenic gene expression were noted. Cbfa1, a necessary transcription factor for osteoblast differentiation, was expressed 33% higher (p= 0.26); Collagen 1, 69% higher (p= 0.05); Osterix, 49% higher (p= 0.001); and BSPII, 54% higher (p= 0.001) than osteoblasts cultured for 72 hours in standard 2D monolayer conditions. When cultured in the presence of collagen alpha2beta1 integrin receptor antibody, 3D aggregates had decreased levels of mineralization as compared with non-treated aggregates. CONCLUSION: RCC enhances osteoblast differentiation using integrin mediated pathways.

Effects of implant surface microtopography on osteoblast gene expression.

AIM: The promotion of osteoblast attachment and differentiation has been evaluated on various implant surfaces. However, the effects of different implant surface properties on gene expression of key osteogenic factors are not fully understood. OBJECTIVE: The objectives of this study were to evaluate how topographical effects on titanium surface alter the expression of bone-related genes and transcription factors. METHODS: Osteoblasts were cultured on titanium disks prepared with a titanium dioxide grit blasting (TiOBlast) or grit blasted and etched with hydrofluoric acid (Osseospeed), grit blasted and etched (SLA-1), or grit blasted, etched and rinsed with N2 protection and stored in isotonic NaCl (SLA-2) commercially pure titanium implant discs. High-density cultures of human mesenchymal pre-osteoblastic cells (HEPM 1486, ATCC) were grown for 72 h and real-time PCR used for quantitative analysis of alkaline phosphatase (ALP), core-binding factor alpha1 (Cbfa1), Osterix, Type I Collagen, Osteocalcin and bone sialoprotein II gene expression. RESULTS: Real-time PCR showed significant (P<0.001) increases in ALP gene expression in osteoblasts grown on SLA-2, relative to all other surfaces. Cbfa1/RUNX-2 gene expression was significantly (P<0.01) increased on Osseospeed and TiOBlast surface as compared with SLA-1 and SLA-2 surfaces. The expression of Osterix had a trend similar to that of Cbfa1. CONCLUSION: In conclusion, implant surface properties may contribute to the regulation of osteoblast differentiation by influencing the level of bone-related genes and transcription factors in human mesenchymal pre-osteoblastic cells.