Primary systemic light chain amyloidosis or immunoglobulin light-chain amyloidosis (AL) is the most common type of systemic amyloidosis.AL is a proteotoxic clonal plasma cell disease, a hematological malignancy, characterised by overproduction of immunoglobulin light chains that form characteristic abnormally folded and aggregated, insoluble fibrillar deposits in various organs, including kidneys, heart, liver, and autonomic and peripheral nerves, etc, these processes lead to organ dysfunction and death. Systemic amyloidosis have various types with different causes, thereby its clinical diagnosis and treatment are more difficult. Recent developments on studies that have significantly aided the management of patients with AL include diagnostic techniques for definitive typing of amyloid deposits by using flow cytometry and immunophenotype analysis. These methods can detect abnormalities of bone marrow plasma cell clones, such as CD38(+), CD138(+), CD56(+), CD19(-) in AL patients. The monitoring abnormal plasma cells with immunoglobulin light chain restriction and abnormal plasma cell phenotypic characteristics contributes to the early diagnosis of AL and detection of minimal residual disease after treatment, which greatly improved AL treatment and prognosis. In this review the diagnosis and typing, clinical characteristics, flow cytometry, immunophenotyping of bone marrow cells, immunoglobulin light chain restriction and phenotypic characteristics of abnormal plasma cells of AL are briefly summarized.
Amyloidosis ; diagnosis ; Bone Marrow Cells ; cytology ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Plasma Cells ; cytology

OBJECTIVETo determine whether human alveolar bone osteoblasts (HABO) attachment to commercial available guided tissue regeneration (GTR) membranes can be enhanced by coating with freshly prepared human platelet-rich plasma (PRP) and platelet poor plasma (PPP).

METHODSHuman osteoblasts established from tissue explants were used at 4 th passage in culture. Human whole blood from healthy subjects was collected and centrifuged twice to produce the PRP fraction and PPP fraction. Double-sided adhesive tape was used to fix 3 mm discs of each membrane and cover-slides to the bottom of a 24-well tissue culture plate. A (GoreTex-ePTFE), B (GoreTex-Resolut) and C (Inion-GTR) membranes were studied. Cover-slides were positive control. Membranes or cover-slides were exposed to PRP, PPP or PBS respectively for 2 hours. Membranes and cover-slides were seeded with osteoblasts (5 x 10(7) cells/L) and allowed to attach for 24 hours. After staining with hematoxylin, the number of attached cells per mm(2) was counted using a light microscope with graticule. The the ultrastructure of osteoblasts attachment to the membranes was observed by scanning electronic microscopy.

RESULTSPRP and PPP-treated membranes significantly enhanced osteoblasts attachment compared to PBS-treated membranes (P < 0.05). There was more osteoblasts attachment in the PRP-treated membranes than in the PPP-treated membranes (P < 0.05). Cover-slides showed more osteoblasts attachment than the three membranes (P < 0.05). B and C membranes showed higher cell attachment than A membranes (P < 0.05). SEM showed that osteoblasts attached to the membranes treated by PRP were spindle and stretched well, and there were platelets, fibrins in a interlaced mesh on the membranes, which appeared to grow in a multiplayer style. The osteoblasts attached to the membranes treated by PPP or PBS were round and partially attached.

CONCLUSIONSPRP and PPP could improve attachment of osteoblasts in the three membranes, and PRP altered and enhanced the way of the attachment to the membranes.

Cell Adhesion ; Cells, Cultured ; Culture Media ; Humans ; Osteoblasts ; cytology ; Platelet-Rich Plasma ; Tooth Socket ; cytology

OBJECTIVETo explore a new method for in vitro expansion of human mesenchymal stem cells (MSC) by using platelet-rich plasma (PRP) in place of fetal calf serum (FCS).

METHODSBone marrow(BM) samples were obtained from the proximal femurs of patients with normal haematopoietic function undergoing total hip arthroplasty. 2 x 10(5)/cm2 BM nucleated cells were seeded in 25 cm2 flasks for MSC cultivation containing one of the 3 mediums: complete Dexter medium with 12.5% FBS and 12.5% horse serum (medium1), alpha-MEM with 10% FCS (medium2) and alpha-MEM with 5% PRP(medium3). At the same time, 1 x 10(6) nucleated BM cells and same amount of nucleated cells from iliac aspirate were seeded in 25 cm2 tissue flasks, colony forming unit-fibroblast (CFU-F) assay.

RESULTSCulture-expanded cells from proximal femurs presented a typical fibroblast-like morphology. Cells were positive for SH2 (CD 105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. On induction, these cells could differentiate into osteoblasts. A significantly higher proliferative capacity of MSCs expanded in medium3 was observed in comparison to those in mediuml or 2 without alteration of the phenotype and the differentiation property. CFU-F assays indicated that bone marrow from the proximal femoral contained significantly more CFU-F than that from iliac aspirate.

CONCLUSIONPlatelet-rich plasma can be used in place of FCS to provide a safer and more effective culture condition to expand MSCs for clinical purpose. The proximal femur BM cells can be obtained in hip surgeries.

Blood Platelets ; Cell Culture Techniques ; methods ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology ; Plasma

OBJECTIVETo observe the effects of platelet-rich plasma (PRP) on inducing mineralization of human gingival fibroblasts (GF), periodontal ligament cells (PDLC) and alveolar bone osteoblasts (AOB).

METHODSHuman whole blood from healthy subjects was collected and PRP was obtained after twice centrifuged. GF, PDLC and AOB established from tissue explants were used at 4th passage in culture. All cells were seeded at density of 4 x 10(7)/L at 6 well tissue culture plate and cultured at standard condition for 96 hours, and then the different media was used. In the experimental group, the cells continued to be cultured in 50 ml/L PRP in DMEM media with mineral solution (10 mmol/L beta-glycerophosphate, 0.2 mmol/L freshly prepared ascorbic acid, 10 nmol/L dexamethasone), in the control group, the cells to be cultured in DMEM media with mineral solution, and in the blank group DMEM media was used. The media was changed every other day until day 30, the cells were dyed with von Kossa. Pictures were taken and analyzed by the image analysis software to semiquantitate the proportion of the positive area of von Kossa dye and attenuation aera of AOB.

RESULTSIn PDLC and AOB, the quantity of mineralization nodule in the experimental group and the control group was more than that in the blank group (P < 0.05), and there were more mineralization nodules in the experimental group than in the control group (P < 0.05), while the amount of mineralization nodules in GF was low in the three groups (P > 0.05). Attenuation of AOB in the experimental group was less than the control group (P < 0.05).

CONCLUSIONSPRP can enhance the quantity of mineralization in PDLC and AOB in vitro and reduce attenuation in AOB, which suggests that PRP may be helpful to periodontal alveolar bone mineralization and regeneration.

Adolescent ; Adult ; Alveolar Process ; cytology ; Cells, Cultured ; Female ; Fibroblasts ; chemistry ; cytology ; Gingiva ; cytology ; Humans ; Male ; Osteoblasts ; chemistry ; cytology ; Periodontal Ligament ; cytology ; Platelet-Rich Plasma ; Young Adult

OBJECTIVETo observe the effect of platelet-rich plasma (PRP) on the proliferation and adipogenic differentiation of human adipose-derived stem cells in vitro.

METHODSHuman adipose-derived stem cells were obtained by enzymatic digestion and PRP was prepared by dual centrifugal method. The ADSCS were interfused with 5%, 10%, and 20% PRP in conditioned culture media, using the untreated cells as the control group. The morphology of the cells were observed and their proliferative ability was detected using XTT colorimetric assay. The adipogenic differentiation ability of the cells was evaluated using oil Red O staining.

RESULTSThe ADSCS treated with PRP showed better morphology with higher density than the control cells. XTT colorimetric assay demonstrated obviously stronger proliferative activity of PRP-treated cells than the control group (P<0.01). Interfusion with PRP caused a significant increase in adipogenic differentiation of the cells as compared to the control cells (P<0.01).

CONCLUSIONPRP treatment produces obvious effects on the proliferation and adipogenic differentiation of human adipose-derived stem cells in vitro.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; Humans ; Lipectomy ; Platelet-Rich Plasma ; Stem Cells ; cytology ; Tissue Engineering ; methods

Humans ; Multiple Myeloma ; blood ; diagnosis ; Plasma Cells ; cytology ; Plasmacytoma ; blood ; diagnosis

OBJECTIVETo construct an injectable tissue-engineered bone graft with fibrin glue (FG), autologous platelet-rich plasma (PRP) and bone marrow stromal cells (BMSCs) cultured in vitro and study its biological characteristics and microscopic structures.

METHODSBMSCs isolated from rabbit iliac bone marrow were culture-expanded in vitro. The injectable tissue-engineered bone constructed from autologous PRP, FG, and BMSCs was cultured in vitro, and its biological characteristics were observed including the time of gel formation, histological features, seed cell survival and microscopic structures.

RESULTSThe constructed injectable tissue-engineered bone began gel formation within 20 to 30 s, and after a week-long culture, the gelatine began to degrade, and numerous well viable fusiform cells could be seen to adhere to the bottom of the Petri dish. Scanning electron microscopy identified globular and olivary cells embedded in the fibrin glue, and numerous small particles could be seen around of the cells.

CONCLUSIONConstruction of an injectable tissue-engineered bone graft with FG, BMSCs and PRP does not require sophisticated techniques and ensures good biological property of the bone graft that can be easily shaped and allow good growth of the seed cells, suggesting great potential of this technique for clinical use.

Bone Marrow Cells ; cytology ; Bone Substitutes ; chemistry ; Cells, Cultured ; Coculture Techniques ; Fibrin Tissue Adhesive ; chemistry ; Platelet-Rich Plasma ; chemistry ; Stromal Cells ; cytology ; Tissue Engineering ; methods

OBJECTIVETo evaluate the effects of different concentrations of platelet-rich plasma (PRP) on human dental pulp stem cells (hDPSC) proliferation and osteogenic differentiation activity so as to provide basis for future application of dental pulp stem cells and PRP in tissue engineering and bone repair therapy.

METHODShDPSC were isolated and cultivated in vitro. Flow cytometric analysis was carried out to test the expression of STRO-1.hDPSC were cultured in various concentrations of PRP (1%, 5%, 10%, 20%). At the 2nd and 6th day 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was tested. Osteogenic differentiation of hDPSC was assessed using alkaline phosphatase (ALP) staining and Alizarin Red staining at the 7th and 14th day.

RESULTSFlow cytometric analysis demonstrated that 14.82% of hDPSC were STRO-1 positive. One percent to 20% PRP showed significant effect of promoting hDPSC proliferation. One percent to 10% PRP showed significant effect of promoting hDPSC osteogenic differentiation.

CONCLUSIONSCertain concentrations of PRP can promote hDPSC proliferate and osteogenic differentiate, and this finding suggests future application of dental pulp stem cells and PRP in bone tissue engineering.

Adolescent ; Adult ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Child ; Dental Pulp ; cytology ; Humans ; Male ; Middle Aged ; Osteogenesis ; Platelet-Rich Plasma ; Stem Cells ; cytology ; Young Adult

OBJECTIVETo investigate whether plasma from patients with systemic lupus erythematosus (SLE) inhibits the suppressive effects of mesenchymal stem cells (MSCs) on lupus B lymphocytes.

METHODSMSCs isolated and expanded from the bone marrow of healthy donors were co-cultured with B cells purified from the peripheral blood of SLE patients in the presence of fetal bovine serum or pooled plasma from SLE patients, and the proliferation and maturation of the B lymphocytes were analyzed.

RESULTSs Co-culture with normal MSCs obviously inhibited the proliferation of lupus B cells and suppressed the maturation of B lymphocytes, which showed lowered expressions of CD27 and CD38. The pooled plasma from SLE patients significantly inhibited the suppressive effects of normal MSCs on B cell proliferation and maturation.

CONCLUSIONPlasma from SLE patients negatively modulates the effects of normal MSCs in suppressing lupus B cell proliferation and maturation to affect the therapeutic effect of MSC transplantation for treatment of SLE. Double filtration plasmapheresis may therefore prove beneficial to enhance the therapeutic effects of MSC transplantation for SLE.

B-Lymphocytes ; pathology ; Cell Proliferation ; Coculture Techniques ; Humans ; Lupus Erythematosus, Systemic ; blood ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; cytology ; Plasma

OBJECTIVE: To determine the effect of pregnancy-associated plasma protein A (PAPP-A) on the function of vascular endothelial cells (VEC). METHODS: Human umbilical vein endothelial cell (HUVEC) line, derived from human umbilical vein, was cultured in vitro with PAPP-A at 0, 50, 100, and 200 ng/mL for 0, 12, 24, and 48 hours, respectively. Nitric oxide (NO) levels and endothlin-1 (ET-1) levels were determined by spectrophotometer and immunehistory. RESULTS: The NO levels of HUVECs in the PAPP-A groups were significantly lower than those in the control group (P<0.05). The ET-1 levels of HUVECs in the PAPP-A groups were significantly higher than those in the control group (P<0.05). The changes were all dose-dependent. CONCLUSION PAPP-A may affect the function of vascular endothelial cells by reducing the secretion of NO and increasing the level of ET-1.
Cell Line ; Endothelial Cells ; cytology ; metabolism ; physiology ; Endothelin-1 ; biosynthesis ; Female ; Humans ; Nitric Oxide ; biosynthesis ; Pregnancy-Associated Plasma Protein-A ; pharmacology ; Umbilical Veins ; cytology ; metabolism