**STATEMENT OF WORK**

1.  **[Objectives]{.underline}**

> **The contractor will provide highly specialized preparation and
> analyses of bone specimens in support of studies in the Skeletal
> Clinical Studies Unit (SCSU) of the Craniofacial and Skeletal Diseases
> Branch (CSDB) of the National Institutes of Dental and Craniofacial
> Research. These will primarily be, but not confined to, specimens from
> studies of patients with hypoparathyroidism.**

2.  **[Approach]{.underline}**

> **The contractor will receive fixed bone specimens obtained from
> patients at the NIH Clinical Center as part of the IRB-approved SCSU
> clinical studies. He will process the specimens in his laboratory, and
> perform the agreed upon analyses. Specimens are property of the US
> government, and except for those parts of the specimens needed for
> future reference by Dr. Dempster, will be returned upon completion of
> the analyses. SCSU investigators and Dr. Dempster and his staff will
> review and analyze the results of his studies in a collaborative
> fashion and prepare figures representative of the findings for
> publication.**

3.  **[Methods]{.underline}**

> **[i. Methods of tissue processing]{.underline}:**
>
> **The bone samples will be processed as previously described (Ref.1).
> The specimen is fixed, dehydrated, and embedded in methyl methacrylate
> without decalcification according to the following schedule:**

**a. Fixative: 10% buffered formaldehyde for 24h;**

**b. 70% ethanol for 6-7days;**

**c. Absolute ethanol for 2 days;**

**d. Toluene for 2 days;**

**e. Activated solution of methyl methacrylate for 4 days;**

**f. Exposure at 32°C, constant temperature, for 3 days.**

> **Undecalcified sections are cut on a Reichert-Jung Polycut microtome
> by 3 levels in a biopsy core with approximately 150 m apart between
> levels. For each level, sections cut at 7 m are stained with
> Goldner\'s trichrome, solochrome and toluidine blue, respectively, for
> routine morphology, and one section cut at 20 m is mounted unstained
> for visualization of tetracycline fluorescence under ultraviolet
> light. Goldner trichrome stain demonstrates bone section in best
> colors, solochrome stain discriminates well between mineralized and
> non-mineralized matrix (osteoid) and toluidine blue stain permits
> visualization of the bone cells, including osteoclasts.**
>
> **[ii. Methods of measurement:]{.underline}**
>
> **Each section will be subjected to histology evaluation and
> histomorphometry, using computerized image analysis, as previously
> described in this laboratory (Ref 1). The computerized image analysis
> system consists of a digitizing tablet, a high resolution color video
> camera, a light microscope with fluorescence system, a color computer
> display and an OsteoMeasure computer software system (version 4.00c)
> for the quantitative analysis of areas, perimeters, distances and
> points.**
>
> **The following parameters will be obtained by measurement on
> cancellous and cortical bone of the biopsy sections:**

# Static Parameters

> **Osteoid perimeter (O.Pm/B.Pm), Osteoid width (O.Wi), Osteoclast
> number (Oc.No/B.Pm), Eroded perimeter (E.Pm/B.Pm), and Wall width of
> cancellous bone packets (W.Wi)**

# Dynamic parameters

> Mineral apposition rate (MAR), Mineralized perimeter (Md.Pm/B.Pm),
> Bone formation rate, tissue level, perimeter referent (BFR/B.Pm),
> Adjusted apposition rate (Aj.AR), Formation period of trabecular bone
> packets (FP), Resorption period (Rs.P), Activation frequency (Ac.f)

# Structural parameters

> **Cancellous bone area (B.Ar/T.Ar), Ratio of bone surface to volume
> (B.Pm/B.Ar), Trabecular width (Tb.Wi), Trabecular number (Tb.N),
> Trabecular separation (Tb.Sp), Cortical width (Ct.Wi), Cortical
> Porosity Density (Ct.Po.N/Ct.Ar ) and Cortical Porosity Area
> (Ct.Po.Ar).**

5.  **[Personnel, Evaluation, and Schedule]{.underline}**

    i.  **[Personnel]{.underline}**

> **Dr. Dempster and his staff are experts in the area of the processing
> and evaluation of human bone biopsy specimens.**
>
> **Personnel will comply with the rules and policies of the NIDCR and
> its staff.**

ii. **[Evaluation]{.underline}**

> **If the contractor's performance is found to be substandard in the
> judgment of the Project Officer, the contractors will initially
> receive notice of the problem from the Project Officer and will be
> required to take corrective measures and to demonstrate improved
> performance.**

iii. **[Schedule]{.underline}**

> **The contractor will work with the investigators and collaborators on
> an as-need basis. It is estimated there will be approximately 10 -- 20
> specimens per year for analysis.**
>
> **5. [Reporting Requirements]{.underline}**
>
> **The contractor will provide bone specimen processing and analysis to
> the SCSU, CSDB, DIR, NIDCR during the period of July 1, 2006 through
> June 3, 2007. The CSDB, in consideration of the foretasted services,
> will pay a computed rate of \$1000 per specimen processed and
> analyzed. He will be required to submit statements to the SCSU
> investigators on an ad hoc basis as the work is completed.**
>
> **Justification**
>
> **Attached is a request for approximately \$20,000.00 to provide a fee
> and travel for the services of Dr. David Dempster. Under this contract
> Dr. Dempster will provide bone specimen processing and analyses to the
> SCSU, CSDB, DIR, NIDCR. His contract will run from July 1, 2006 --
> June 30, 2007.**
>
> **Dr. Dempster is a nationally-recognized expert in bone biology, and
> human bone specimen processing and analyses. These are skills that we
> have been unable to secure on the NIH campus. He has numerous
> publications in the field and has collaborated with several NIH
> researchers in the past as well as currently.**
>
> **Appointment of Dr. Dempster is of critical importance to assure that
> the rare clinical specimens that are acquired as part of SCSU clinical
> studies are appropriately prepared and studied. Lack of expertise will
> not only jeopardize the quality of the project and validity of the
> data, but will result in research subjects having gone through
> invasive research procedures for no benefit to the field. For these
> reasons it is requested that this request be processed as quickly as
> possible.**

**Reference**

1\. Dempster DW, Cosman F, Kurland ES, Zhou H, Nieves J, Woelfert L,
Shane E, Plavetic K, Muller R, Bilezikian J, Lindsay R 2001 Effects of
daily treatment with parathyroid hormone on bone microarchitecture and
turnover in patients with osteoporosis: A paired biopsy study. J Bone
Miner Res 16(10): 1846-1853.