Histomorphometric assessment of bone necrosis produced by two cryosurgery protocols using liquid nitrogen: an experimental study on rat femurs

Abstract

ObjectiveThe aim of this study was to evaluate the effects of liquid nitrogen cryosurgery on the femoral diaphysis of rats. Material and MethodsThe femoral diaphyses of 42 Wistar rats were exposed to three local and sequential applications of liquid nitrogen for 1 or 2 min, intercalated with periods of 5 min of passive thawing. The animals were sacrificed after 1, 2, 4 and 12 weeks and the specimens obtained were processed and analyzed histomorphometrically. ResultsThe depth and extent of peak bone necrosis were 124.509 μm and 2087.094 μm for the 1-min protocol, respectively, and 436.424 μm and 12046.426 μm for the 2-min protocol. Peak necrosis was observed in the second experimental week with both cryotherapy protocols. ConclusionsThe present results indicate that the 2-min protocol produced more marked bone necrosis than the 1-min protocol. Although our results cannot be entirely extrapolated to clinical practice, they contribute to the understanding of the behavior of bone tissue submitted to different cycles of liquid nitrogen freezing and may serve as a basis for new studies.

Histological and Immunological Evaluation of the Osteogenic Effects of Compact Bone–Delivered Stem Cell on Spongiosis Bone in the Rat Zygomatic Arch Defect Model

“…27 As a result of this method, cells suffer cytotoxic damage. [27][28][29] In our study, the method used by Costa et al 19 in rat trabecular and spongiosis bones was used. According to this protocol, the zygomatic bone is immersed in liquid nitrogen for 1 minute and passively thawed for 5 minutes, and this cycle is repeated 3 times.…”

“…Most studies investigating the effect of stem cells on bone defects have focused on the femur bone. 13,14,19 Because it is a large trabecular bone, it is easier to create defects. In our study, however, a zygomatic bone was used.…”

“…To obtain an acellular scaffold from bone tissue, we applied a cryotherapy procedure, which Costa et al 19 stated is ideal for cancellous bones. The zygomatic bone was immersed in liquid nitrogen for 1 minute, and passive thawing (incubation at room temperature) was performed for 5 minutes.…”

The Effect of Liquid Nitrogen on Bone Graft Survival

“…5,15,17 In the evaluation of experimental and clinical studies, the duration of freezing in the liquid nitrogen is variable and there is no consensus between surgeons. There are some reports that recommend holding the bone tissue in liquid nitrogen just for a few minutes 7,12,13,16,19 whereas in some reports the duration of freezing was prolonged up to 30 minutes. 6,15,18,20 The crystallization that causes the damage to the bone cells rises with the duration of freezing, but increased freezing time also generates irreversible weakening in the bone tissue.…”

Evaluation of different therapeutic Carnoy's formulations on hard human tissues: A Raman microspectroscopy, microhardness, and scanning electron microscopy study

“…(Abou Neel et al, 2016) Conservative treatment of these jaw lesions can lead to high recurrence rates; however, a radical approach usually is followed by severe functional and aesthetic impairments (Costa et al, 2010;Pogrel, 2015). Adjunct therapies allied to surgical treatments, such as the use of Carnoy's solution or liquid nitrogen as a cryosurgical agent, have been widely used since the 1980s, aiming to improve clinical outcomes (Costa et al, 2011). Voorsmit, Stoelinga andvan Haelst in 1981 (Voorsmit et al, 1981) were the first investigators that used Carnoy's solution in intraosseous-related jaw lesions, recommending the use of this cauterizing agent in the treatment of odontogenic keratocysts, as this lesion shows important infiltration patterns and high rates of recurrence (Voorsmit et al, 1981).…”