Chondrocyte Proliferation

PCNA Staining

PCNA Staining

Neonatal bone sections are common tissues to study chondrocyte proliferation. Immunohistochemical PCNA labeling in paraffin sections is fairly straight-forward. BIOQUANT’s thresholding tools automatically identify PCNA+ cells based on stain color. Ideally, it's best to keep the counter-stain light to avoid interference with the DAB chromagen.

Proliferation Zone Boundary

Irregular region of interest

Define the contours of the proliferation zone using BIOQUANT’s Irregular Region of Interest tool. BIOQUANT ensures that only chondrocytes within the proliferation zone are counted, and records the area data needed to normalize the number of proliferating cells by the size of the zone.

Detecting PCNA+ Cells

Detection is based on chromagen color and cell morphology. BIOQUANT's Measure Cells tool identifies PCNA+ chondrocytes using the DAB chromagen. Then, labeled cells are passed through size and shape filters to exclude non-specific staining.

BIOQUANT’s Measure Cells tool also filters out cells that have already been counted as we profiles that intersect the edge of the proliferation zone.

Detecting PCNA- Cells

Then Measure Cells identifies PCNA- chondrocytes in the same field of view. These cells are also passed through size and shape filters. Additionally, BIOQUANT knows that PCNA- cells are never PCNA+. Finally, BIOQUANT will not count cells partially in the field of view or that have been previously counted.

Automation

auotmatic roi & batch Scripts

Using the Automatic ROI tool and a batch script, BIOQUANT can sequence through every field of view to count cells and still ensure that only cells within the proliferation boundary are counted.

BIOQUANT automatically computes:

Pr.Zn.Ar (Proliferation Zone Area)
PCNA+ Ch.N (PCNA+ Chondrocyte Number)
PCNA- Ch.N (PCNA- Chondrocyte Number)
Tt.Ch.N (Total Chondrocyte Number)
PI (Proliferation Index)
PCNA+ Ch.N / Pr.Z.Ar

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Recent Citations of BIOQUANT in Cell Proliferation

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osteoblast, teriparatide, Gli1+, MMPs, proliferation, COP

Gli1+ progenitors mediate bone anabolic function of teriparatide via Hh and Igf signaling

osteoblast, teriparatide, Gli1+, MMPs, proliferation, COP

diabetes, implant, proliferation, rabbit, CS/PK, cranial bone defect, osteointegration

Incorporation of nanosized calcium silicate improved osteointegration of polyetheretherketone under diabetic conditions

diabetes, implant, proliferation, rabbit, CS/PK, cranial bone defect, osteointegration

Diabetes can impair osteoblastic functions and negatively interfere with osteointegration at the bone/implant interface. Previously, we prepared a nanosized calcium silicate (CS) incorporated-polyetheretherketone (PK) biocomposite (CS/PK) and found that the CS/PK composite exhibited enhanced osteoblast functions in vitro and osteointegration in vivo, but its bioperformance under diabetic conditions remained elusive.

diabetes, implant, proliferation, rabbit, CS/PK, cranial bone defect, osteointegration

allograft, PET mesh, autograft, proliferation

Cell and Tissue Response to Polyethylene Terephthalate Mesh Containing Bone Allograft in Vitro and in Vivo

allograft, PET mesh, autograft, proliferation

halofuginone, metastasis, anti-angiogenic, proliferation, TGF

Halofuginone inhibits TGF-β/BMP signaling and in combination with zoledronic acid enhances inhibition of breast cancer bone metastasis

halofuginone, metastasis, anti-angiogenic, proliferation, TGF

More efficient therapies that target multiple molecular mechanisms are needed for the treatment of incurable bone metastases. Halofuginone is a plant alkaloid-derivative with antiangiogenic and antiproliferative effects. Here we demonstrate that halofuginone is an effective therapy for the treatment of bone metastases, through multiple actions that include inhibition of TGFβ and BMP-signaling.

halofuginone, metastasis, anti-angiogenic, proliferation, TGF

rat, anabolic, peptin, proliferation, alanine scan

Structure activity relationship study on the peptide hormone preptin, a novel bone-anabolic agent for the treatment of osteoporosis

rat, anabolic, peptin, proliferation, alanine scan

Preptin is a 34-residue pancreatic hormone shown to be anabolic to bone in vitro and in vivo. The bone activity of preptin resides within the (1-16) N-terminal fragment. Due to its peptidic nature, the truncated fragment of preptin is enzymatically unstable; however it provides an attractive framework for the creation of stable analogues using various peptidomimetic techniques.

rat, anabolic, peptin, proliferation, alanine scan

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