J Korean Neurosurg Soc.  2017 Jul;60(4):397-403. 10.3340/jkns.2017.0101.004.

Osteoblast and Bacterial Culture from Cryopreserved Skull Flap after Craniectomy: Laboratory Study

• Cho TG
• Kang SH
• Cho YJ
• Choi HJ
• Jeon JP
• Yang JS

• KMID: 2387877
• DOI: http://doi.org/10.3340/jkns.2017.0101.004

Abstract

OBJECTIVE
Cranioplasty using a cryopreserved skull flap is a wide spread practice. The most well-known complications of cranioplasty are postoperative surgical infections and bone flap resorption. In order to find biological evidence of cryopreserved cranioplasty, we investigated microorganism contamination of cryopreserved skulls and cultured osteoblasts from cryopreserved skulls.
METHODS
Cryopreserved skull flaps of expired patients stored in a bone bank were used. Cryopreserved skulls were packaged in a plastic bag and wrapped with cotton cloth twice. After being crushed by a hammer, cancellous bone between the inner and outer table was obtained. The cancellous bone chips were thawed in a water bath of 30°C rapidly. After this, osteoblast culture and general microorganism culture were executed. Osteoblast cultures were done for 3 weeks. Microorganism cultures were done for 72 hours.
RESULTS
A total of 47 cryopreserved skull flaps obtained from craniectomy was enrolled. Of the sample, 11 people were women, and the average age of patients was 55.8 years. Twenty four people had traumatic brain injuries, and 23 people had vascular diseases. Among the patients with traumatic brain injuries, two had fracture compound comminuted depressed. The duration of cryopreservation was, on average, 83.2 months (9 to 161 months). No cultured osteoblast was observed. No microorganisms were cultured.
CONCLUSION
In this study, neither microorganisms nor osteoblasts were cultured. The biological validity of cryopreserved skulls cranioplasty was considered low. However, the usage of cryopreserved skulls for cranioplasty is worthy of further investigation in the aspect of cost-effectiveness and risk-benefit of post-cranioplasty infection.

Keyword

Decompressive craniectomy; Cranioplst; Cryopreservation; Osteoblast; Cell culture techniques; Bacterial infections

MeSH Terms

Bacterial Infections
Baths
Bone Banks
Brain Injuries
Cell Culture Techniques
Cryopreservation
Decompressive Craniectomy
Female
Humans
Osteoblasts*
Plastics
Skull*
Vascular Diseases
Water
Plastics
Water

Figure

• 

  Fig. 1 Osteoblast extraction method from cryopreserved skull. A: The cryopreserved skull flap packages were crushed by hammer and bone fragments were used. Using sterile surgical instrument, cancellous bone between inner and outer table was obtained. B: Low power field polarization microscopic examintaion 21 days after culture showed abundant spindle-like cells around bone chips (×40). C: High power field microscopic examination with alkaline phosphatase staining showed purple colored stained cell as described in the Materials and Methods sections (ALP staining, ×200).

• 

  Fig. 2 Schematic figure of bone fusion process after craniotomy. A: Inflammatory phase. The bone flap is surrounded by blood and inflammatory response is initiated. (Top to bottom) Capillaries from surrounding bone, dura and periosteum infiltrate to the transplanted bone. As granulation tissue proliferated, capillaries invade the transplanted bone flap. Through the capillary, primitive progenitor cell migrated and bone remodeling occurred. If this functional contact between the transplanted flap and surrounding bone is poor, re-inserted bone flap would be in ischemic necrosis, and be absorbed. B: Callus formation phase. Cartilage and fibrous tissue is laid down and make new lamellar bone, which is remodeled with osteoclast-osteoblast coupling activity to strong bone fusion.

Reference

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